Split type space biological culture device and system

Abstract

The invention belongs to the technical field of space biological culture, and provides a split type space biological culture device and system.The device comprises a first substrate, a chip, a second substrate and a PDMS sealing film, the second substrate is provided with a chip groove, the chip is arranged in the chip groove, the chip comprises a first sealing layer, a middle layer and a second sealing layer which are sequentially arranged from top to bottom, and the first sealing layer is arranged on the middle layer; the middle layer is provided with a cavity, a liquid inlet flow channel and a liquid outlet flow channel, the fluid contact surfaces of the cavity, the liquid inlet flow channel and the liquid outlet flow channel are all made of PDMS materials, the PDMS sealing film is arranged between the first sealing layer and the first substrate, covers the cavity and avoids a liquid inlet connector and a liquid outlet connector corresponding to the liquid inlet flow channel and the liquid outlet flow channel, and the first substrate, the second substrate and the chip are of a split structure. According to the scheme, large-capacity biological culture can be achieved, the risk of biocompatibility of materials is eliminated through split processing of the device body and the chip, the use cost is reduced, the culture chip is of a multi-layer structure, and the mechanical strength and the capacity of the cavity are guaranteed.

Description

Technical Field

[0001] This application belongs to the field of space biological culture technology, specifically relating to a split-type space biological culture device and system. Background Technology

[0002] With the development of space life sciences, research on the reproductive system function and developmental patterns of organisms in space is becoming increasingly crucial. In such research, specialized devices capable of supporting the development and culture of microfluidic living organisms such as embryos and cells under the unique conditions of space microgravity are core equipment for ensuring the smooth conduct of experiments. These devices must not only adapt to the limited payload of space launches and the resource constraints of on-orbit experiments, but also provide a stable and controllable culture environment for living organisms to meet the stringent requirements of space life science research. Summary of the Invention

[0003] In order to overcome the shortcomings of the prior art, this application provides a split-type space biological culture device.

[0004] This application provides a split-type space biological culture device, wherein the biological entity includes a living biological entity with a three-dimensional spatial structure, and the device includes: a first substrate, a chip, a second substrate, and a PDMS sealing film; The second substrate has a chip slot, and the chip is disposed in the chip slot; The chip includes a first sealing layer, an intermediate layer, and a second sealing layer arranged sequentially from top to bottom. The intermediate layer has a chamber, a liquid inlet channel, and a liquid outlet channel. The fluid contact surfaces of the chamber, the liquid inlet channel, and the liquid outlet channel are all made of PDMS material. The opening of the chamber is exposed to the first sealing layer. The liquid inlet channel communicates with the chamber and is used to allow liquid to enter the chamber. The liquid outlet channel communicates with the chamber and is used to allow liquid to exit the chamber. The chip is also provided with an inlet port connecting the inlet channel and the surface of the first sealing layer, and an outlet port connecting the outlet channel and the surface of the first sealing layer, for the purpose of supplying nutrients to the organism and discharging waste liquid. The PDMS sealing film is disposed between the first sealing layer and the first substrate. The PDMS sealing film covers the chamber opening on the surface of the first sealing layer and avoids the inlet port and outlet port corresponding to the inlet channel and outlet channel. The first substrate, the second substrate, the PDMS sealing film, and the chip are separate structures.

[0005] Optionally, the intermediate layer includes a first intermediate layer and a second intermediate layer bonded to each other, the liquid outlet channel is etched on the surface of the first intermediate layer facing the first sealing layer, and the liquid inlet channel is etched on the surface of the second intermediate layer facing the second sealing layer. The first intermediate layer is provided with multiple first sub-cavities, and the second intermediate layer is provided with second sub-cavities that correspond one-to-one with the first sub-cavities; The first sub-cavity is bonded to the corresponding second sub-cavity to form the chamber.

[0006] Optionally, the inlet channel is connected to the bottom of the plurality of second sub-cavities, and the outlet channel is connected to the top of the plurality of first sub-cavities, forming a fluid path that flows from bottom to top.

[0007] Optionally, the chip is fabricated through the following steps: A pattern of the liquid inlet channel is etched into the surface of the second intermediate layer and recessed towards the surface of the second sealing layer. A through hole is machined on the second intermediate layer to form a second compartment. A through hole is machined at the corresponding position of the liquid inlet interface to complete the structural forming of the liquid inlet interface on the second intermediate layer. A pattern of the liquid outlet channel is etched into the surface of the first intermediate layer facing the first sealing layer. A through hole is processed on the first intermediate layer to form a first cavity. Through holes are processed in the first intermediate layer and the second intermediate layer respectively corresponding to the position of the liquid inlet interface, so as to complete the structural forming of the liquid inlet interface on the first intermediate layer. Align and bond the surface of the first intermediate layer facing away from the first sealing layer with the surface of the second intermediate layer facing away from the second sealing layer to form an intermediate layer assembly; process through holes at the position corresponding to the chamber of the first sealing layer to form a chamber opening; process through holes at the position corresponding to the liquid inlet to form a liquid inlet; process through holes at the position corresponding to the outlet of the liquid outlet channel to form a liquid outlet. Align the lower surface of the first sealing layer facing the first intermediate layer with the upper surface of the intermediate layer assembly to ensure precise alignment of the liquid outlet with the outlet of the liquid flow channel, the chamber opening with the chamber, and the liquid inlet with the inlet of the liquid flow channel, and then complete the bonding between the first sealing layer and the intermediate layer assembly; seal the liquid flow channel pattern on the upper surface of the first intermediate layer with the first sealing layer to form a sealed liquid flow channel. Finally, the lower surface of the intermediate layer component is aligned and bonded to the upper surface of the second sealing layer. The liquid inlet channel pattern on the lower surface of the second intermediate layer is sealed by the second sealing layer to form a closed liquid inlet channel, thus completing the overall fabrication of the chip.

[0008] Optionally, the chamber includes at least one chamber group, with multiple chambers in each chamber group arranged in a straight line. Each chamber group includes multiple culture chambers and an observation chamber. The observation chamber is located in the middle of the corresponding group. The multiple culture chambers are symmetrically distributed with the observation chamber as the center. The number of chambers in each group is an odd number. Correspondingly, each liquid inlet channel corresponds one-to-one with the chamber group; each liquid inlet channel includes a multi-level tree-like branch structure, and the last level tree-like branch structure is connected to the culture chamber in the chamber group corresponding to the liquid inlet channel. Each liquid inlet channel also includes a curved pipe, which is connected to the observation chamber in the chamber group corresponding to the liquid inlet channel.

[0009] Optionally, the liquid outlet channels correspond one-to-one with the chamber groups, each liquid inlet channel corresponds to a different liquid inlet port, and each liquid outlet channel is connected to the same liquid outlet port.

[0010] Optionally, the liquid outlet channel integrates a Tesla valve structure, which consists of multiple teardrop-shaped chambers connected in series to prevent waste liquid backflow.

[0011] Optionally, the first substrate has a first connector screw hole and a second connector screw hole, the first connector screw hole is provided with a liquid inlet connector, and the second connector screw hole is provided with a liquid outlet connector. A steel pin is provided on the lower surface of the first substrate corresponding to the screw holes of the first connector and the screw holes of the second connector. One end of the steel pin is sealed and fitted with the liquid inlet connector or the liquid outlet connector, and the other end passes through the clearance hole of the PDMS sealing film and is sealed and connected with the liquid inlet interface or the liquid outlet interface.

[0012] Optionally, both the inlet and outlet connectors are Peek connectors, the steel needle is a hollow round tube, and the insertion depth of the steel needle into the inlet or outlet interface is 1-2 mm.

[0013] Optionally, the first substrate and the second substrate are provided with a plurality of sets of fixing screw holes, and fixing screws are provided in the fixing screw holes to lock and fix the first substrate, PDMS sealing film, chip and second substrate.

[0014] The present invention also proposes a split-type space biological culture system, which includes the split-type space biological culture device as described above.

[0015] This application achieves functional separation of the chamber and flow channel interface through a separate design of the first substrate, second substrate, PDMS sealing film, and chip, combined with the flexible clamping seal of the PDMS sealing film. The chamber is fully enclosed for protection, while the inlet and outlet interfaces remain open and connected, balancing sealing safety and ease of fluid handling, and providing a structural foundation for long-term stable operation in space experimental environments. The use of full PDMS contact path encapsulation of the chamber and fluid channels completely isolates toxic substances that may leach from external mounting components, creating a safe and clean culture environment for living biological entities in space. Simultaneously, the separate space biological culture device adopts a separate structure design for the device body and chip, allowing users to conduct different experiments simply by replacing the chip as needed, without having to remanufacture the entire device body. This significantly reduces the overall processing and maintenance costs of the device, balancing experimental safety and economy. Attached Figure Description

[0016] To more clearly illustrate the technical solutions of the embodiments of this application, the accompanying drawings used in the embodiments will be briefly introduced below. It should be understood that the following drawings only show some embodiments of this application and should not be regarded as a limitation of the scope. For those skilled in the art, other related drawings can be obtained based on these drawings without creative effort.

[0017] Figure 1 This is a schematic diagram of the structure of the split-type space biological culture device provided in Example 1; Figure 2 for Figure 1 A schematic diagram of the structure of the device after the fixing components have been removed; Figure 3 This is a schematic diagram of another optional structure for a split-type space biological culture device; Figure 4 for Figure 1 A schematic diagram of the structure of the chip; Figure 5 This is a schematic diagram of the front structure of the chip; Figure 6 This is a schematic diagram of the back structure of the chip; Figure 7 This is a schematic diagram of the cavity structure; Figure 8 This is a schematic diagram of the structure of the first substrate.

[0018] Figure label: 1-First substrate; 2-Chip; 3-Second substrate; 4-Chip slot; 6-PDMS sealing film; 11-First connector screw hole; 12-Second connector screw hole; 13-First fixing screw hole; 14-Second fixing screw hole; 21-First sealing layer; 22-First intermediate layer; 23-Second intermediate layer; 24-Second sealing layer; 25-Cavity; 26-Outlet port; 27-First inlet port; 28-Second inlet port; 51-First inlet connector; 52-Second inlet connector; 53-Outlet connector; 54-First inlet needle; 55-Second inlet needle; 56-Outlet needle; 57-Fixing screw; 221-Outlet channel; 231-Inlet channel; 251-First set of chambers; 252-Second set of chambers; 253-Cultivation chamber; 254-Observation chamber; 2311-Bent tubing. Detailed Implementation

[0019] The technical solutions of the embodiments of this application will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only a part of the embodiments of this application, and not all of the embodiments. Based on the embodiments of this application, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of this application.

[0020] In the description of this application, it should be noted that the terms "vertical", "up", "down", "horizontal", etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings. They are only for the convenience of describing this application and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limitations on this application.

[0021] In the description of this application, it should also be noted that, unless otherwise expressly specified and limited, the terms "set," "install," "connect," and "link" should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral connection; they can refer to a mechanical connection or an electrical connection; they can refer to a direct connection or an indirect connection through an intermediate medium; and they can refer to the internal connection of two components. Those skilled in the art can understand the specific meaning of the above terms in this application according to the specific circumstances.

[0022] Existing space culture devices are generally made of polycarbonate (PC). During the perfusion and transfer process, it is difficult to avoid direct contact between the external culture medium and the PC material. Substances leached from the PC material can seep into the chamber through fluid exchange. If the living organisms are sensitive to their environment, this could pose a safety hazard to their survival and development. In addition, existing space culture devices are mostly integrated designs, which not only result in complex overall structures leading to high processing difficulties and manufacturing costs, but also have the drawback of requiring the entire device to be scrapped if there is local contamination or damage. Even if only a local area is contaminated or slightly damaged, the entire space culture device needs to be replaced, which significantly increases the cost of use and maintenance and seriously restricts the economy and continuity of space life science experiments.

[0023] In view of this, this application provides a split-type space biological culture device, which fundamentally solves the biocompatibility problem of traditional devices. It employs a full PDMS contact path to encapsulate the chamber and fluid channels, completely isolating toxic substances that may leach from external components, thus creating a safe and clean culture environment for living biological entities in space. Furthermore, the split-type space biological culture device adopts a separate structure design for the main body and the chip, allowing users to conduct different experiments simply by replacing the chip as needed, without having to remanufacture the main body. This significantly reduces the overall processing, use, and maintenance costs of the device, balancing experimental safety and economy.

[0024] The modular space biological culture device provided in this application is suitable for the culture of live biological entities in a space environment. Live biological entities refer to living objects with a three-dimensional spatial structure, including but not limited to: embryos, cells, three-dimensional tissues, and organoids. Cells refer to independent single cells or cell populations; three-dimensional tissues refer to cell-matrix complexes without organ function; organoids refer to three-dimensional structures that simulate organ function; and embryos refer to early living organisms developed from fertilized eggs. The size range of this live biological entity in all dimensions is 1µm-10mm, preferably 10µm-1mm. The modular space biological culture device adopts a compact design, adaptable to the limited installation space within a spacecraft. The arrangement of each component has been precisely optimized, integrating complete culture, liquid inlet, liquid outlet, and observation functions within a very small volume.

[0025] Please see Figures 1-3 The split-type space biological culture device provided in this application embodiment may include: a first substrate 1, a chip 2, and a second substrate 3. The second substrate 3 has a chip slot 4 for accommodating the chip 2.

[0026] The chip slot 4 can be a rectangular groove, with its length and width dimensions matching those of the chip 2. The depth of the chip slot 4 is precisely designed to match the thickness of the chip 2. That is, the depth of the chip slot 4 is not less than the thickness of the chip 2, so that after the chip 2 is placed in the chip slot 4, the upper surface of the chip 2 is basically flush with or slightly convex to the upper surface of the second substrate 3. As a support base for the chip 2, the chip slot 4 plays a positioning role for the chip 2, providing stable support for the chip 2, ensuring that the chip 2 will not shift during assembly, and preventing displacement of the chip 2 under the vibration of space launch or the microgravity environment of space, thus ensuring the overall structural strength of the device. At the same time, it also ensures that the clamping force can be evenly applied to the chip 2 when it is subsequently locked with the first substrate 1, avoiding clamping failure or liquid leakage caused by excessive slot depth or chip height, and fully meeting the stringent requirements of space experiments.

[0027] For example, the first substrate 1 and the second substrate 3 can be made of acrylic or PC material. Acrylic material has high light transmittance, which is convenient for observation, while PC material has better strength and can withstand more severe mechanical environments. Both materials have good processing performance and biocompatibility, and will not cause pollution to the culture environment. Moreover, the thickness design can ensure the depth of the chip groove 4 to meet the installation requirements of the chip 2.

[0028] The first substrate 1, chip 2, PDMS sealing film 6, and second substrate 3 are manufactured separately and then assembled into a single structure. The separate design facilitates the precision manufacturing of each component, and in actual use, users can replace the chip as needed to carry out different experiments without having to remake the main body of the device (first substrate 1 and second substrate 3), which greatly reduces the overall manufacturing and maintenance costs of the device, while taking into account both experimental safety and economy.

[0029] See Figure 4 As shown, chip 2 has a four-layer composite structure, including a first sealing layer 21, a first intermediate layer 22, a second intermediate layer 23, and a second sealing layer 24 arranged sequentially from top to bottom. The four-layer structure is formed into a single chip body through a bonding process. The overall thickness is adapted to the depth of the chip slot 4, ensuring the overall structural strength of the split-type space biological culture device and adapting to the space experimental environment.

[0030] Please refer to the following: Figure 5 and Figure 6As shown, the first sealing layer 21 is made of PDMS material. Its front side (upper surface) is used to adhere the PDMS sealing film 6, and its back side (lower surface) is tightly bonded to the first intermediate layer 22. The first sealing layer 21 has liquid inlet ports (first liquid inlet port 27 and second liquid inlet port 28) penetrating the first intermediate layer 22 and the second intermediate layer 23, and a liquid outlet port 26 penetrating the first sealing layer 21, used to supply nutrients to the organism and discharge waste liquid. The first liquid inlet port 27, the second liquid inlet port 28, and the liquid outlet port 26 all avoid the projection area of ​​the chamber 25 on the first sealing layer 21. That is, the first liquid inlet port 27, the second liquid inlet port 28, and the liquid outlet port 26 only serve as through ports for fluid channels and do not directly contact the chamber 25.

[0031] Both the first intermediate layer 22 and the second intermediate layer 23 can be made of PDMS material. The surface of the first intermediate layer 22 facing the first sealing layer 21 is etched with a pattern of liquid outlet channel 221. The first intermediate layer and the first sealing layer 21 form the liquid outlet channel 221. The surface of the second intermediate layer 23 facing the second sealing layer 24 is etched with a pattern of liquid inlet channel 231. The second intermediate layer 23 and the second sealing layer 24 form the liquid inlet channel 231. The liquid inlet interface connects the liquid inlet channel 231 and the surface of the first sealing layer 21. The liquid outlet interface 26 connects the liquid outlet channel 221 and the surface of the first sealing layer 21.

[0032] Furthermore, the first intermediate layer 22 has multiple first cavities penetrating through it, and the second intermediate layer 23 has multiple second cavities penetrating it. The positions, numbers, sizes, and shapes of the first and second cavities are matched. For each first cavity, there is a second cavity whose projection on the horizontal plane coincides with its own. The first cavity and the second cavity whose projection on the horizontal plane coincides with its own are considered as corresponding first and second cavities. The first sealing layer forms through holes corresponding to the positions of the first cavities as openings of the first cavities on the chip surface. After the openings of the first sealing layer 21, the first cavities, and the corresponding second cavities are coaxially aligned and bonded, and then bonded to the second sealing layer, a complete cavity 25 is formed, wherein the second sealing layer serves as the bottom of the cavity. The sum of the thickness of the first sealing layer 21 and the depths of the first and second cavities is the total depth of the cavity 25. The opening of chamber 25 is exposed to the first sealing layer 21. The top of chamber 25 is connected to the liquid outlet channel 221 through the liquid outlet channel branch of the first intermediate layer 22, and the bottom of chamber 25 is connected to the liquid inlet channel 231 through the liquid inlet channel branch of the second intermediate layer 23, realizing a bottom-inlet and top-outlet fluid path, effectively avoiding the generation of air bubbles in the chamber and ensuring the stability of the culture environment for live biological entities.

[0033] In existing split-type space biological culture devices, the inlet and outlet channels connecting the chambers are usually at the same height. Through extensive experiments, the inventors of this application have discovered that the existing structure is unsuitable for culturing living organisms with three-dimensional spatial structures in space. This is because culturing living organisms with three-dimensional spatial structures requires the culture chamber to have a certain height. When the inlet and outlet channels are at the same height, some liquid will flow into the chamber from the inlet channel and then directly flow out from the outlet channel, without filling the chamber completely. This phenomenon is even more pronounced in microgravity or weightlessness, resulting in air bubbles remaining in the chamber after liquid inflow. The presence of these air bubbles not only occupies the effective culture space of the living organisms, interfering with their growth and nutrient exchange, but may also cause damage to the living organisms due to the localized impact force generated by bubble bursting, seriously affecting the stability and data accuracy of space culture experiments.

[0034] This application embodiment addresses the need for culturing live biological entities in a microgravity environment in space. Combining the three-dimensional characteristics of live biological entities such as embryos, cells, three-dimensional tissues, and organoids, the chamber 43 is designed as a blind hole structure with a certain height. It also adopts a three-dimensional flow channel layout and a bottom-in, top-out fluid perfusion structure, which solves the problems of bubble retention and uneven perfusion in traditional devices. During the initial liquid introduction, the culture medium enters from the bottom of chamber 25 through the inlet channel 231. As the liquid gradually rises, it pushes the gas in the chamber upwards, allowing the bubbles to be smoothly discharged from the top of chamber 25 through the outlet channel 221. During the liquid replacement operation, new culture medium is also injected from the bottom of chamber 25. The rising liquid flow replaces the old culture medium in the chamber layer by layer, and the waste liquid is discharged from the outlet channel 221 at the top. This design allows for buoyancy-assisted gas removal and liquid replacement under normal gravity conditions, while under microgravity conditions, surface tension and the spatial layout of the channels guide fluid movement, effectively forming a stable fluid replacement process, reducing the risk of bubble retention, and ensuring that each chamber 25 receives sufficient and consistent culture medium exchange, greatly improving the reliability and repeatability of space biological culture experiments.

[0035] The second sealing layer 24 is made of high-transmittance glass material. Its upper surface is tightly bonded to the lower surface of the second intermediate layer 23. It is used to seal the liquid inlet channel 231 and provide a light-transmitting observation window, ensuring that the microscope can clearly observe the growth status of the living biological entity in the chamber through the second sealing layer 24. At the same time, the flatness of the glass material can avoid the problem of uneven refractive index when the PDMS material transmits light, thus improving the observation accuracy.

[0036] During chip fabrication, a pattern for the liquid inlet channel is first etched into the lower surface of the second intermediate layer 23 (the surface facing the second sealing layer 24). Then, through-holes are processed on the second intermediate layer 23 to form a second sub-cavity. Through-holes are also processed at corresponding positions of the first liquid inlet interface 27 and the second liquid inlet interface 28 to complete the structural formation of the first liquid inlet interface 27 and the second liquid inlet interface on the second intermediate layer 23. A pattern for the liquid outlet channel is etched into the upper surface of the first intermediate layer 22 (the surface facing the first sealing layer 21). Then, through-holes are processed on the first intermediate layer 22 to form a first sub-cavity. Through-holes are also processed in the first intermediate layer 22 and the second intermediate layer 23 at positions corresponding to the first liquid inlet interface 27 and the second liquid inlet interface 28 to complete the structural formation of the first liquid inlet interface 27 and the second liquid inlet interface 28 on the first intermediate layer 22. After the above processing is completed, the lower surface of the first intermediate layer 22 (the surface facing away from the first sealing layer 21) and the upper surface of the second intermediate layer 23 (the surface facing away from the second sealing layer 24) are aligned and bonded to form an intermediate layer assembly. The first and second cavities on this intermediate layer assembly are bonded to form a chamber 25. It should be noted that the through-hole processing of the first intermediate layer 22 and the second intermediate layer 23 is completed before bonding. This avoids damage to the bonding surface caused by drilling after bonding, and also avoids the problem of drilling not being able to penetrate due to the intermediate layer assembly being too thick after bonding. Then, through-holes are processed at the positions of the chambers corresponding to the first sealing layer 21 to form the chamber openings of 10 chambers; through-holes are processed at the corresponding positions of the first liquid inlet 27 and the second liquid inlet 28 to form liquid inlet interfaces; and through-holes are processed at the corresponding positions of the outlet of the liquid outlet channel 221 to form the liquid outlet interface 26. Align the lower surface of the first sealing layer 21 (the surface facing the first intermediate layer 22) with the upper surface of the intermediate layer assembly (the upper surface of the first intermediate layer 22), ensuring precise alignment of the outlet port 26 with the outlet channel 221, the chamber opening with the chamber, and the inlet port with the inlet channel. Then, bond the first sealing layer 21 to the intermediate layer assembly. The first sealing layer 21 seals the outlet channel pattern on the upper surface of the first intermediate layer 22, forming a sealed outlet channel 221. Finally, align and bond the lower surface of the intermediate layer assembly (the lower surface of the second intermediate layer 23) with the upper surface of the second sealing layer 24 (the surface facing the second intermediate layer 23). The second sealing layer 24 seals the inlet channel pattern on the lower surface of the second intermediate layer 23, forming a sealed inlet channel 231, thus completing the overall fabrication of the chip 2.

[0037] The PDMS sealing film 6 is made of PDMS material and has a dense, non-porous sheet structure with dimensions consistent with the upper surface dimensions of the first sealing layer 21. The PDMS sealing film 6 is laid on the upper surface of the first sealing layer 21, located between the first sealing layer 21 and the first substrate 1, and is used to completely cover the openings of each chamber on the first sealing layer 21 to prevent external contaminants from entering.

[0038] For example, please see Figures 5-6 The surface of the first sealing layer 21 is rectangular. The chamber opening can be located at the center of the surface of the first sealing layer. The first liquid inlet 27 is located near the middle of one of the long sides of the rectangle. The second liquid inlet 28 is located near the middle of the other long side of the rectangle. The liquid outlet 26 is located near the middle of one of the short sides of the rectangle. The PDMS sealing film 6 can cover the center of the surface of the first sealing layer so that the PDMS sealing film 6 just covers the chamber opening without covering the liquid inlet or liquid outlet.

[0039] Therefore, the size of the PDMS sealing membrane 6 can be designed so that it precisely covers the chamber opening without covering the inlet or outlet ports. Alternatively, a larger PDMS sealing membrane 6 can be used, with clearance holes of the same diameter opened on the PDMS sealing membrane 6 at positions corresponding to the first inlet port 27, the second inlet port 28, and the outlet port 26. The clearance holes are precisely aligned with each port to ensure that the flow of fluid is not affected, thus blocking the chamber opening and avoiding the inlet and outlet ports.

[0040] See Figure 7 As shown, in an optional embodiment, considering the limited payload and experimental resources within the space capsule, in order to conduct multiple parallel control experiments simultaneously in a limited space, the multiple chambers 25 of chip 2 can be divided into at least one chamber group. The following description uses two groups of chambers (i.e., the first group of chambers 251 and the second group of chambers 252) as an example.

[0041] Furthermore, the multiple chambers are set up independently. Independent setup means that each chamber has its own independent inlet port. Specifically, the first chamber 251 is equipped with a first inlet port 27, and the second chamber 252 is equipped with a second inlet port 28. Equipping each chamber with an independent inlet port ensures that the flow rate and concentration of the culture medium in each chamber can be independently controlled, avoiding cross-interference caused by interconnection in traditional parallel flow channels, and ensuring the accuracy and uniqueness of experimental conditions for different experimental groups. Each chamber can also be equipped with an independent outlet port 26, achieving independent control of the entire inlet and outlet process; provided that back diffusion interference does not occur between the culture media, multiple chambers can also share a single outlet port 26 to simplify the device structure, reduce the number of ports, and lower the device weight.

[0042] Based on the independent setup of the multiple chambers mentioned above, this embodiment can be used to perfuse biological reagents or culture media with different components and concentrations to meet the differentiated needs of parallel experiments in different groups. This allows multiple independent control experiments to be carried out simultaneously in the same device, such as comparative studies of different drug dosages and different culture environment parameters. This improves the operational efficiency of spatial experiments and the ability to process samples in parallel, effectively overcoming the technical limitations of the existing technology where a single device is used to carry out a single group of experiments and the resource utilization rate is low.

[0043] Furthermore, each group of chambers can adopt an odd number of designs, specifically including multiple culture chambers 253 and one observation chamber 254, wherein the observation chamber 254 is located in the middle of the corresponding group, and the culture chambers 253 are symmetrically distributed with the observation chamber 254 as the center.

[0044] This embodiment features a 5-chamber structure with 4 culture chambers 253 and 1 observation chamber 254. Adjacent chambers within the same group are spaced 5 mm apart, while chambers in different groups are spaced 10 mm apart. This design ensures uniform fluid distribution among the chambers, provides a convenient central location for observation operations, and achieves efficient adaptation between the culture space and observation functions.

[0045] The culture chamber 253, with a diameter of 4 mm, a height of 8 mm, a cross-sectional curvature of 1.7785 rad, and an arc length of 2.4 mm, serves as the core carrier for biological culture. Its large radial dimensions and volume provide ample growth space for living organisms, meeting their developmental and metabolic needs throughout the culture cycle. All experimental data are taken from culture chamber 253, eliminating the need for full-field observation and ensuring the smooth progress of the live organism culture process. The observation chamber 254, with a diameter of 3 mm, a height of 8 mm, a cross-sectional curvature of 2.95135 rad, and an arc length of 1.6 mm, is precisely sized to match the field of view of a high-powered microscope, allowing for the acquisition of complete images in a single operation. It is specifically designed for detailed morphological observation, dynamic development tracking, or monitoring of specific targets, and its independent design from the culture chambers prevents interference with the main culture environment during observation operations.

[0046] Furthermore, the multiple chambers in the first group of chambers 251 can be linearly arranged along a first straight line, and the multiple chambers in the second group of chambers 252 can be linearly arranged along a second straight line, with the first straight line parallel to the second straight line. This layout facilitates the flow channel connection process and provides a regular field of view for observation operations.

[0047] See Figure 5As shown, the inlet channel 231 is located in the edge region of the upper surface of the second intermediate layer 23. The inlet channel 231 includes one or more levels of tree-like branching structure. Each parent channel in the inlet channel 231 is divided into at least two sub-channels of the same geometric size. The last sub-channel is connected to the corresponding chamber to distribute the culture medium evenly to each chamber. Each inlet channel 231 may also include a curved tube 2311, which is connected to the observation chamber in the chamber group corresponding to the inlet channel 231.

[0048] A tree-like branching structure is a network system that efficiently divides a main channel, which acts as the trunk, into multiple smaller terminal channels, which act as the leaves. The core feature of a tree-like branching structure is its hierarchical and decreasing structure, which can branch from a large main channel into more and finer sub-channels, eventually covering all chambers. Following physical and geometric optimization principles, the tree-like branching structure can minimize flow resistance and maximize distribution efficiency, ensuring the uniform distribution of the culture medium.

[0049] For example, the liquid inlet channel 231 may include a first liquid inlet channel and a second liquid inlet channel that are independent of each other. Each liquid inlet channel includes a three-level tree-like branch structure, including a first-level tree-like channel, a second-level tree-like channel and a third-level tree-like channel. The ends of the three-level tree-like channels are respectively connected to the bottom of each chamber. The starting ends of the first-level tree-like channels are precisely aligned with the first liquid inlet interface 27 and the second liquid inlet interface 28 on the first sealing layer 21, respectively, and are connected to the external fluid channel through the through opening of the first sealing layer 21.

[0050] Furthermore, two inlet channels 231 are symmetrically distributed on both sides of the chip 2, with a tree-like branching structure extending towards the center of the chip. The ends of the three-level tree-like channels are perpendicular to the bottom center of each chamber, ensuring that the culture medium flows in uniformly from the bottom center of the chamber. The inlet channels 231 are directly connected to the bottom of each chamber, realizing a bottom-entry fluid path design. Specifically, the last sub-channel of the first inlet channel is connected to each chamber in the first group of chambers 251, and the last sub-channel of the second inlet channel is connected to each chamber in the second group of chambers 252.

[0051] To accommodate the layout of the two sets of chambers described above, the first liquid inlet channel includes one primary channel, two secondary channels, and four tertiary channels. The primary channels connect the primary liquid inlet 27 to each of the secondary channels. Two secondary channels on either side each connect to two tertiary channels, and each tertiary channel connects to a culture chamber within the first set of chambers 251. The first liquid inlet channel also includes a curved conduit 2311 located between the two secondary channels. The curved conduit 2311 connects to the observation chamber within the first set of chambers 251 and is connected to the primary channels. The flow path of the curved conduit can be precisely designed so that the flow resistance encountered by the liquid flowing from the primary liquid inlet through the curved conduit 2311 into the observation chamber is essentially the same as the flow resistance encountered by the liquid flowing from the primary liquid inlet through the secondary and tertiary channels into the culture chamber. The second inlet channel includes one primary channel, two secondary channels, and four tertiary channels. The primary channels connect the second inlet port 28 to each of the secondary channels. Two secondary channels on either side each connect to two tertiary channels. Each tertiary channel connects to a culture chamber 253 within the second set of chambers 252. The second inlet channel also includes a curved conduit 2311 located between the two secondary channels, connecting to an observation chamber 254 within the second set of chambers 252. This curved conduit 2311 is connected to the primary channel. The flow path of the curved conduit 2311 can be precisely designed so that the flow resistance encountered by the liquid flowing from the second inlet port 28 through the curved conduit 2311 into the observation chamber 254 is essentially the same as the flow resistance encountered by the liquid flowing from the second inlet port 28 through the secondary and tertiary channels into the culture chambers 253. This channel design achieves uniform distribution of the culture medium to each chamber, balancing channel fabrication feasibility and fluid uniformity.

[0052] See Figure 4 As shown, the liquid outlet channel 221 is located at the edge region of the upper surface of the first intermediate layer 22. The liquid outlet channel 221 includes interconnected branch channels and a main channel. The end of the main channel (liquid outlet end) is located in the middle region of the upper surface of the first intermediate layer 22, precisely aligned with the liquid outlet interface 26 of the first sealing layer 21. The branch channels extend horizontally from the top of each chamber to the main channel, which is arranged along the length of the chip 2. The ends of the branch channels of the liquid outlet channel 221 are directly connected to the top of each chamber, realizing the upward fluid path design.

[0053] Furthermore, the liquid outlet channel 221 is provided with a Tesla valve structure 2211 to prevent waste liquid from flowing back into each chamber; specifically, the Tesla valve structure 2211 can be integrated into the middle section of the main channel of the liquid outlet channel 221.

[0054] A Tesla valve is a one-way valve without moving parts. Unlike traditional valves that rely on valve discs, balls, or diaphragms, Tesla valves guide fluid purely through their special geometry, making the resistance to fluid flow in one direction much smaller than in the opposite direction.

[0055] In the above embodiment, by integrating a Tesla valve structure consisting of multiple teardrop-shaped chambers connected in series on the liquid outlet channel 221, it is possible to effectively prevent waste liquid from flowing back into the chamber. Even if each chamber 25 is connected to the same liquid outlet interface, it is possible to avoid cross-contamination of waste liquid in each chamber 25.

[0056] See Figures 4-5 As shown, the first liquid inlet 27, the second liquid inlet 28, and the liquid outlet 26 are all circular through holes penetrating the first sealing layer 21. The through holes on the first sealing layer 21, the first intermediate layer 22, the second intermediate layer 23, and the second sealing layer 24 can be formed using drilling tools such as a hole punch, or they can be prepared by molding or laser drilling processes to ensure the accuracy of the hole diameter and the integrity of the structure.

[0057] For example, the first liquid inlet 27 and the second liquid inlet 28 are symmetrically distributed at both ends of the width direction of the chip 2, respectively located at the center points of the two long sides of the chip 2. They are located on the upper surface of the first sealing layer 21 and precisely correspond to... Figure 8 The first liquid inlet 27 and the second liquid inlet 28 are located directly below the first liquid inlet needle 54 and the second liquid inlet needle 55 on the lower surface of the first substrate 1. The lower ends of the first liquid inlet interface 27 and the second liquid inlet interface 28 are directly connected to the liquid inlet channel 231 of the second intermediate layer 23. The upper end of the first liquid inlet interface 27 is tightly fitted with the lower end of the first liquid inlet needle 54, and the upper end of the second liquid inlet interface 28 is tightly fitted with the lower end of the second liquid inlet needle 55, forming a gapless fluid channel. At the same time, the projection of the first liquid inlet interface 27 on the XY plane completely coincides with the inlet of the first-stage tree-like flow channel of the first liquid inlet channel, and the projection of the second liquid inlet interface 28 on the XY plane completely coincides with the inlet of the first-stage tree-like flow channel of the second liquid inlet channel, ensuring that the fluid is introduced without deviation.

[0058] The liquid outlet 26 is located on the upper surface of the first sealing layer 21, corresponding to... Figure 8 The liquid outlet 26 is located directly below the liquid outlet needle 56 on the lower surface of the first substrate 1. For example, the liquid outlet 26 is located at the center of one short side of the chip. The lower end of the liquid outlet 26 is directly connected to the end of the main channel of the liquid outlet flow channel of the first intermediate layer 22, and the upper end is tightly fitted to the lower end of the liquid outlet needle 56, forming a gapless fluid channel. Moreover, its projection on the XY plane completely coincides with the end of the main channel of the liquid outlet flow channel 221, ensuring smooth discharge of waste liquid.

[0059] It is worth noting that the liquid inlet is inserted through the first sealing layer 21, the first intermediate layer 22 and the second intermediate layer 23, thereby connecting the liquid inlet channel 231 located between the second intermediate layer 23 and the second sealing layer 24. The liquid outlet 26 is inserted only through the first sealing layer 21, thereby connecting the liquid outlet channel 221 located between the first sealing layer 21 and the first intermediate layer 22. This structural design not only meets the fluid conduction requirements, but also simplifies the processing flow.

[0060] Optionally, the first inlet connector 51, the second inlet connector 52, and the outlet connector 53 all use Peek connectors (made of polyetheretherketone). The connector is cylindrical in shape and has a through-flow fluid channel inside, the diameter of which is precisely matched with the inner diameter of the steel needle. The upper end of the connector has an external thread for connecting to the external infusion tubing, and the lower end has a tapered interface for a tight fit with the steel needle. Peek material has excellent biocompatibility, corrosion resistance, and high temperature resistance, and will not release toxic substances that contaminate the culture environment. At the same time, the threaded connection ensures reliable sealing with the external infusion tubing.

[0061] See Figure 1 As shown, the first substrate 2 has two first connector screw holes 11 (corresponding to two liquid inlet connectors) and one second connector screw hole 12 (corresponding to one liquid outlet connector). All screw holes are internally threaded, with specifications matching the external threads of the Peek connector. Specifically, the first liquid inlet connector 51 is installed in one of the first connector screw holes 11, the second liquid inlet connector 52 is installed in the other first connector screw hole 11, and the liquid outlet connector 53 is installed in the second connector screw hole 12. The axis of each connector is completely aligned with the axis of the corresponding steel needle.

[0062] Furthermore, the projection of the inlet connector on the horizontal plane coincides with the end of the inlet channel furthest from the chamber, and the projection of the outlet connector on the horizontal plane coincides with the end of the outlet channel furthest from the chamber. This precise alignment design provides a structural basis for efficient fluid conduction. Simultaneously, the first inlet needle 54 passes through the top of the first sealing layer 21 and is inserted into the corresponding first inlet port 27; the second inlet needle 55 passes through the top of the first sealing layer 21 and is inserted into the corresponding second inlet port 28; and the outlet needle 56 passes through the top of the first sealing layer 21 and is inserted into the corresponding outlet port 26. The user only needs to align the first inlet needle 54 with the first inlet port 27, the second inlet needle 55 with the second inlet port 28, and the outlet needle 56 with the outlet port 26 above the chip 2 to quickly achieve stable communication between the inlet connector, inlet port, and inlet channel, as well as between the outlet connector, outlet port, and outlet channel.

[0063] The tapered interface at the lower end of the liquid inlet connector is inserted into the interior of the upper end of the first liquid inlet steel needle 54 and the second liquid inlet steel needle 55, and the tapered interface at the lower end of the liquid outlet connector 53 is inserted into the interior of the upper end of the liquid outlet steel needle 56. The tapered structure achieves an interference fit, effectively preventing fluid leakage. At the same time, the liquid inlet connector and the liquid outlet connector are tightened and fixed to the connector screw hole of the first base plate 1 through external threads, ensuring that they will not loosen in the microgravity environment of space.

[0064] The first liquid inlet needle 54, the second liquid inlet needle 55, and the liquid outlet needle 56 are all hollow round tubes. The surface of the needles is polished, with no burrs or scratches, ensuring smooth fluid flow without damaging the PDMS sealing film 6 and the chip interface.

[0065] The first liquid inlet needle 54, the second liquid inlet needle 55, and the liquid outlet needle 56 are all vertically fixed to the lower surface of the first substrate 1. The first liquid inlet needle 54 and the second liquid inlet needle 55 are directly opposite the first connector screw hole 11, and the liquid outlet needle 56 is directly opposite the second connector screw hole 12, extending from the lower surface of the first substrate 1 towards the second substrate 3. The needles are fixed to the first substrate 1 by an interference fit, ensuring that the needles remain secure and do not tilt after fixing, thus guaranteeing structural stability.

[0066] The core function of the steel needle is to act as a flow-guiding bridge between the connector and the opening of chip 2, enabling precise fluid conduction. Its upper end is interference-fitted with the tapered interface of the Peek connector, and its lower end penetrates the clearance hole of the PDMS sealing film 6 and is inserted into the corresponding interface of chip 2 at a depth of 1-2mm, ensuring a tight fit with the flow channel inlet without gaps, thereby preventing fluid leakage or the generation of air bubbles.

[0067] During fluid transport, the culture medium is introduced from the first inlet connector, flows sequentially through the first inlet needle 54, the first inlet port 27, and the inlet channel 231 before reaching the chamber 25, and / or, the culture medium is introduced from the second inlet connector, flows sequentially through the second inlet needle 55, the second inlet port 28, and the inlet channel 231 before reaching the chamber 25; while the waste liquid in the chamber 25 flows sequentially through the outlet channel 221, the outlet port 26, the outlet needle 56, and the outlet connector 53, and is finally discharged from the outlet port 26, forming a complete and stable fluid circulation path.

[0068] like Figures 1-3As shown, the first substrate 1 and the second substrate 3 have multiple sets of fixing screw holes. Four first fixing screw holes 13 are respectively located at the four outer corners of the two substrates for installing tightening screws. Four second fixing screw holes 14 are located on the outer area of ​​the chip slot 4, with four on each of the first substrate 1 and the second substrate 3, arranged in a rectangular pattern around the chip slot 4, for installing M3 screws. The tightening screws are Phillips head screws, and the M3 screws are countersunk Phillips head screws. The screws are made of stainless steel with a galvanized surface to effectively prevent rust and meet the requirements of space environments.

[0069] After the screw passes through the fixing screw hole of the first substrate 1, it is threadedly connected to the fixing screw hole of the second substrate 3. During the tightening process, the axial force of the screw causes the first substrate 1 to press down on the PDMS sealing film 6 and the chip 2, so that the lower end of the chip 2 is tightly attached to the bottom of the chip groove 4 of the second substrate 3, forming a stable assembly structure.

[0070] This split-type space biological culture device adopts an independent and precise fluid communication design. Specifically, the first liquid inlet 27 is sealed and connected to the first liquid inlet channel through the first liquid inlet connector 51, and the end of the first liquid inlet channel is connected to each chamber in the first chamber group. The second liquid inlet 28 is sealed and connected to the second liquid inlet channel through the second liquid inlet connector 52, and the end of the second liquid inlet channel is connected to each chamber in the second chamber group. The liquid outlet 26 is sealed and connected to the liquid outlet channel 221 through the liquid outlet connector 53, and the waste liquid from both chambers flows into the liquid outlet channel for unified discharge.

[0071] This design achieves complete independence for the two liquid inlet systems, allowing for the separate injection of culture media with different compositions and concentrations into the two chambers. This enables simultaneous control experiments within a single device, significantly improving experimental efficiency and the accuracy of data comparison. Furthermore, the complete interconnected path of "connector-steel needle-interface-flow channel-chamber," combined with the sealing structure of each component, ensures the airtightness and stability of fluid transmission, avoiding cross-contamination between different flow channels. At the same time, the centralized design of the liquid outlet simplifies the device structure, adapting to the resource constraints and miniaturization requirements of the space environment.

[0072] The first substrate 1, as the core load-bearing component, has first connector screw holes 11 and 12 adapted to the Peek connector, and first fixing screw holes 13 and 14 matched with the fixing screws 57. On its lower surface, corresponding to the positions of the connector screw holes, a first liquid inlet steel needle 54, a second liquid inlet steel needle 55, and a liquid outlet steel needle 56 are vertically fixed. The steel needles and the substrate are securely connected through an interference fit, without loosening or tilting. Through locking with the second substrate 3, the first substrate 1 can apply uniform pressure to the chip 2 and the PDMS sealing film 6, achieving reliable fixation and sealing, and also provides stable support for the connection between external pipelines and the steel needles. Its compact structural layout makes full use of limited space, ensuring the overall miniaturization of the device and adapting to the stringent installation space constraints within the spacecraft. This split-type space biological culture device adopts an independent and precise fluid communication design, forming a complete closed-loop communication path of "connector-steel needle-interface-flow channel-chamber". Specifically, the external culture medium is first introduced into the device through a Peek connector with external threads, and then introduced into a vertically set steel needle through the internal through channel of the connector. The lower end of the steel needle is precisely inserted into the first liquid inlet 27 or the second liquid inlet 28 of the chip 2, forming a gapless seal with the interface through an interference fit. Subsequently, the culture medium flows into the liquid inlet channel 231 of the second intermediate layer 23 through the interface and is introduced into the first set of chambers 251 or the second set of chambers 252. The waste liquid after culture flows out from the top of the chamber and flows into the liquid outlet channel 221 of the first intermediate layer 22. After being treated by the integrated Tesla valve structure to prevent backflow, it enters the liquid outlet steel needle 56 through the liquid outlet 26 and is finally discharged from the device through the liquid outlet connector 53. This design not only achieves relative independence between the two liquid inlet systems, allowing for the separate infusion of culture media with different compositions and concentrations into the two chambers, but also enables simultaneous control experiments within a single device, significantly improving experimental efficiency and the accuracy of data comparison. Furthermore, the precise alignment and sealing of each component ensures the airtightness and stability of fluid transmission, preventing cross-contamination between different channels. At the same time, the centralized design of the liquid outlet channel and the optimized layout of the tree-like channel further simplify the device structure, balance the complexity of the channels with the feasibility of fabrication, and fully adapt to the resource constraints, miniaturization requirements, and high-throughput culture requirements of the space environment.

[0073] In actual use, place the second substrate 3 flat in a laminar flow hood. Remove the chip 2 from its aseptic packaging and place it in the laminar flow hood. Place the chip 2 into the chip slot 4 of the second substrate 3, ensuring the inlet and outlet ports of the chip are facing correctly. Using a micropipette, add live biological entities (embryos, cells, etc., possibly along with some culture medium) sequentially to each of the 10 chamber openings on the first sealing layer 21. Then, lay a PDMS sealing film 6 on the upper surface of the first sealing layer 21 of the chip 2, ensuring the PDMS sealing film 6 completely covers the openings of each culture chamber. Align the first substrate 1 with the second substrate 3, inserting the inlet needles 54 and 55 on the lower surface of the first substrate 1 into the inlet port of the chip 2 (when the PDMS sealing film 6 has clearance holes, the inlet needles 54 and 55 also need to pass through the clearance holes on the PDMS sealing film 6 corresponding to the inlet ports), and inserting the outlet needle 56 on the lower surface of the first substrate 1 into the chip. 2. Inside the liquid outlet interface (when the PDMS sealing membrane 6 is provided with a clearance hole, the liquid outlet steel needle 56 also needs to pass through the clearance hole on the PDMS sealing membrane 6 corresponding to the liquid outlet interface); install the four tightening screws and the four M3 screws into the corresponding fixing screw holes in sequence, and tighten the screws in a diagonal order with a torque wrench to ensure that each component fits tightly and is not loose; install the first liquid inlet connector 51, the second liquid inlet connector 52, and the liquid outlet connector 53 into the corresponding connector screw holes respectively, and tighten them until the lower end of the connector and the upper end of the steel needle fit tightly without gaps.

[0074] This application has the following technical effects: The device employs a full PDMS contact path, with the chamber and all fluid contact parts constructed from PDMS material. A PDMS sealing membrane 6 isolates the chamber, preventing precipitates from external components and fundamentally eliminating biocompatibility risks associated with the material, thus ensuring the safe culture of living biological entities such as embryos and cells. The device features a separate design for the main body and the chip, allowing users to replace the chip as needed to conduct different experiments without having to remanufacture the main body, significantly reducing the overall processing, use, and maintenance costs of the device, and balancing experimental safety and economy.

[0075] The main body of the device and chip 2 are fabricated separately. The main body of the device can be reused for a long time, while chip 2 can be replaced as needed. The intermediate layer is divided into a first intermediate layer 22 and a second intermediate layer 23. Flow channels and cavities are processed on the surfaces of the two layers respectively. The flow channels rely on the surface of the intermediate layer to form a closed structure, which avoids the scrapping of the entire device due to local contamination or changes in experimental requirements, and greatly reduces the cost of use and maintenance. It also reduces the difficulty of processing complex flow channels and deep cavities at the same time, reduces processing costs, and solves the processing problem caused by excessive PDMS film thickness.

[0076] Chip 2 adopts a 4-layer structure, with the middle layer being thicker than the upper and lower sealing layers; the liquid inlet channel 231, liquid outlet channel 221, and chambers are distributed in layers within chip 2, and the connectors and screws are concentrated at the edge and outer side of the substrate; the depth of chip groove 4 is not less than the thickness of chip 2 to ensure the mechanical strength of chip 2 and meet the structural stability requirements in the space environment; it achieves a compact design of the device, makes full use of limited space, adapts to the installation constraints in the space capsule, and integrates complete culture, liquid inlet, liquid outlet, and observation functions in a very small volume; it maintains the overall structural strength of the device and avoids component displacement.

[0077] The first substrate 1 and the first substrate 3 are tightly locked together by multiple fixing screws 57 (four tightening screws + four M3 screws); the PDMS sealing film 6 precisely covers all chambers and avoids the liquid inlet and liquid outlet; the liquid inlet connector, the first liquid inlet steel needle 54, the second liquid inlet steel needle 55, the liquid inlet interface, the liquid inlet channel 231, and the liquid outlet connector 53, the liquid outlet steel needle 56, the liquid outlet interface 26, and the liquid outlet channel 221 are precisely aligned to form a sealed communication path.

[0078] The inlet channel 231 adopts a single-level or multi-level tree-like branch structure to achieve equal distribution of culture medium to all chambers, ensuring uniform perfusion and balancing the complexity of the channel, processing feasibility and fluid uniformity; through the backflow prevention design without power or moving parts, it avoids backflow of waste liquid and cross-contamination between chambers, protecting the cleanliness and stability of the culture environment; it effectively avoids the generation of air bubbles in the chambers and ensures the stability of the culture environment.

[0079] By setting up a completely independent liquid inlet system for each group of chambers (first liquid inlet connector 51, first liquid inlet channel, first liquid inlet interface 27, and second liquid inlet connector 52, second liquid inlet channel, and second liquid inlet interface 28), culture media of different components or concentrations can be injected into different groups of chambers, allowing multiple different experiments to be carried out simultaneously and forming a control, thereby improving experimental efficiency and data comparison accuracy, and maximizing the use of space resources.

[0080] Each chamber is arranged linearly along a parallel straight line, with the central chamber serving as the observation chamber. The diameter of the observation chamber matches the microscope's field of view, facilitating flow channel connectivity and observation feedback. This allows for clear and real-time observation of the biological sample's condition within the chamber, improving observation convenience and data accuracy.

[0081] The second sealing layer 24 is a glass sealing layer, and the first substrate 1 is transparent. The connector is set above the first substrate 1, and the steel needle is directly inserted into the chip to achieve communication, which reduces the difficulty of processing and assembly and makes it easier for astronauts to connect pipelines and change liquids in the space environment.

[0082] This application also provides a split-type space biological culture system, including: The split-type space biological culture device as described in any of the embodiments above.

[0083] In other embodiments, the split-type space biological culture system may further include: The drive module is connected to the multi-stage inlet channel 231 and outlet channel 221 of the split-type space biological culture device, providing controlled fluid power. The heating module is used to heat the split-type space biological culture device to maintain a constant temperature suitable for biological growth, so as to monitor the biological development process. The imaging module is used for real-time imaging of the split-type space biological culture device.

[0084] Specifically, the drive module includes peristaltic pumps or micro-injection pumps, but is not limited to these. As a fluid power source, the drive module provides stable power for the fluid circulation of the split-type space biological culture device. Its output pressure can be precisely matched to the pressure resistance performance of the split-type space biological culture device, and it can be adapted to the serial liquid channel design of the chip to ensure that the culture medium is evenly perfused into each chamber and avoid the generation of air bubbles.

[0085] Specifically, the heating module includes, but is not limited to, thin-film heaters or ambient temperature control chambers. The heating module provides stable temperature support for the split-type space biological culture device, simulating the environment required for biological development and avoiding the impact of extreme temperature changes in space on biological growth.

[0086] Specifically, the imaging module includes, but is not limited to, automated microscopes or CCD cameras. The imaging module performs real-time imaging of organisms within the modular space-based biological culture device.

[0087] The split-type space biological culture system may also include a test solution module. This module includes culture medium bags, waste liquid bags, and fixative bags, which are connected to the split-type space biological culture device in a sealed manner via interface components. It is responsible for providing the culture medium, experimental drugs, and fixative for immobilizing live biological entities required for biological culture, while simultaneously recovering the waste liquid after culture. The entire fluid transmission process is kept airtight to prevent fluid leakage or contamination in the space environment.

[0088] In summary, this application proposes a split-type space biological culture device and system. The proposed solution can realize large-capacity biological culture. By processing the main body of the device and the culture chip separately, the risk of biocompatibility of materials is eliminated and the cost of use is reduced. The culture chip uses a multi-layer structure to ensure mechanical strength and chamber capacity.

[0089] Those skilled in the art will understand that the modules described above can be implemented using general-purpose computing systems. They can be centralized on a single computing system or distributed across a network of multiple computing systems. Optionally, they can be implemented using computer-executable program code, allowing them to be stored in a storage system for execution by the computing system. Alternatively, they can be fabricated as separate integrated circuit modules, or multiple modules or steps can be fabricated as a single integrated circuit module. Thus, this application is not limited to any particular combination of hardware and software.

[0090] Note that the above description is merely a preferred embodiment and the technical principles employed in this application. Those skilled in the art will understand that this application is not limited to the specific embodiments described herein, and various obvious changes, readjustments, and substitutions can be made without departing from the scope of protection of this application. Therefore, although this application has been described in detail through the above embodiments, this application is not limited to the above embodiments. Many other equivalent embodiments may be included without departing from the concept of this application, and the scope of this application is determined by the scope of the appended claims.

[0091] The above disclosures are only a few specific implementation scenarios of this application. However, this application is not limited to these. Any variations that can be conceived by those skilled in the art should fall within the protection scope of this application.

[0092] The above description is only a preferred embodiment of this application and is not intended to limit this application. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of this application should be included within the protection scope of this application.

Claims

1. A split-type space biological culture device, characterized in that, The organism includes a living biological entity with a three-dimensional spatial structure, and the device includes: a first substrate, a chip, a second substrate, and a PDMS sealing film; The second substrate has a chip slot, and the chip is disposed in the chip slot; The chip includes a first sealing layer, an intermediate layer, and a second sealing layer arranged sequentially from top to bottom. The intermediate layer has a chamber, a liquid inlet channel, and a liquid outlet channel. The fluid contact surfaces of the chamber, the liquid inlet channel, and the liquid outlet channel are all made of PDMS material. The opening of the chamber is exposed to the first sealing layer. The liquid inlet channel communicates with the chamber and is used to allow liquid to enter the chamber. The liquid outlet channel communicates with the chamber and is used to allow liquid to exit the chamber. The chip is also provided with an inlet port connecting the inlet channel and the surface of the first sealing layer, and an outlet port connecting the outlet channel and the surface of the first sealing layer, for the purpose of supplying nutrients to the organism and discharging waste liquid. The PDMS sealing film is disposed between the first sealing layer and the first substrate. The PDMS sealing film covers the chamber opening on the surface of the first sealing layer and avoids the inlet port and the outlet port. The first substrate, the second substrate, the PDMS sealing film, and the chip are separate structures.

2. The split-type space biological culture device according to claim 1, characterized in that, The intermediate layer includes a first intermediate layer and a second intermediate layer bonded to each other. The liquid outlet channel is etched on the surface of the first intermediate layer facing the first sealing layer, and the liquid inlet channel is etched on the surface of the second intermediate layer facing the second sealing layer. The first intermediate layer is provided with multiple first sub-cavities, and the second intermediate layer is provided with second sub-cavities that correspond one-to-one with the first sub-cavities; The first sub-cavity is bonded to the corresponding second sub-cavity to form the chamber.

3. The split-type space biological culture device according to claim 2, characterized in that, The inlet channel is connected to the bottom of the plurality of second sub-cavities, and the outlet channel is connected to the top of the plurality of first sub-cavities, forming a fluid path that flows from bottom to top.

4. The split-type space biological culture device according to claim 3, characterized in that, The chip is prepared through the following steps: A pattern of the liquid inlet channel is etched into the surface of the second intermediate layer and recessed towards the surface of the second sealing layer. A through hole is machined on the second intermediate layer to form a second compartment. A through hole is machined at the corresponding position of the liquid inlet interface to complete the structural forming of the liquid inlet interface on the second intermediate layer. A pattern of the liquid outlet channel is etched into the surface of the first intermediate layer facing the first sealing layer. A through hole is processed on the first intermediate layer to form a first cavity. Through holes are processed in the first intermediate layer and the second intermediate layer respectively corresponding to the position of the liquid inlet interface, so as to complete the structural forming of the liquid inlet interface on the first intermediate layer. Align and bond the surface of the first intermediate layer facing away from the first sealing layer with the surface of the second intermediate layer facing away from the second sealing layer to form an intermediate layer assembly; process through holes at the position corresponding to the chamber of the first sealing layer to form a chamber opening; process through holes at the position corresponding to the liquid inlet to form a liquid inlet; process through holes at the position corresponding to the outlet of the liquid outlet channel to form a liquid outlet. Align the lower surface of the first sealing layer facing the first intermediate layer with the upper surface of the intermediate layer assembly to ensure precise alignment of the liquid outlet with the outlet of the liquid flow channel, the chamber opening with the chamber, and the liquid inlet with the inlet of the liquid flow channel, and then complete the bonding between the first sealing layer and the intermediate layer assembly; seal the liquid flow channel pattern on the upper surface of the first intermediate layer with the first sealing layer to form a sealed liquid flow channel. Finally, the lower surface of the intermediate layer component is aligned and bonded to the upper surface of the second sealing layer. The liquid inlet channel pattern on the lower surface of the second intermediate layer is sealed by the second sealing layer to form a closed liquid inlet channel, thus completing the overall fabrication of the chip.

5. The split-type space biological culture device according to claim 4, characterized in that, The chamber includes at least one chamber group, with multiple chambers in each chamber group arranged in a straight line. Each chamber group includes multiple culture chambers and an observation chamber. The observation chamber is located in the middle of the corresponding group. The multiple culture chambers are symmetrically distributed with the observation chamber as the center. The number of chambers in each group is an odd number. Correspondingly, each liquid inlet channel corresponds one-to-one with the chamber group; each liquid inlet channel includes a multi-level tree-like branch structure, and the last level tree-like branch structure is connected to the culture chamber in the chamber group corresponding to the liquid inlet channel. Each liquid inlet channel also includes a curved pipe, which is connected to the observation chamber in the chamber group corresponding to the liquid inlet channel.

6. The split-type space biological culture device according to claim 4, characterized in that, Each liquid outlet channel corresponds to one of the chamber groups, each liquid inlet channel corresponds to a different liquid inlet port, and each liquid outlet channel is connected to the same liquid outlet port.

7. The split-type space biological culture device according to claim 2, characterized in that, The liquid outlet channel integrates a Tesla valve structure, which consists of multiple teardrop-shaped chambers connected in series to prevent waste liquid backflow.

8. The split-type space biological culture device according to claim 1, characterized in that, The first substrate has a first connector screw hole and a second connector screw hole. The first connector screw hole is provided with a liquid inlet connector and the second connector screw hole is provided with a liquid outlet connector. A steel pin is provided on the lower surface of the first substrate corresponding to the screw holes of the first connector and the screw holes of the second connector. One end of the steel pin is sealed to the liquid inlet connector or the liquid outlet connector, and the other end is sealed to the liquid inlet interface or the liquid outlet interface.

9. The split-type space biological culture device according to claim 8, characterized in that, Both the inlet and outlet connectors are Peek connectors, the steel needle is a hollow round tube, and the insertion depth of the steel needle into the inlet or outlet interface is 1-2 mm.

10. The split-type space biological culture device according to claim 8, characterized in that, The first substrate and the second substrate have multiple sets of fixing screw holes, and fixing screws are provided in the fixing screw holes. The fixing screws lock and fix the first substrate, PDMS sealing film, chip and second substrate.

11. A split-type space biological culture system, characterized in that, Includes the split-type space biological culture device as described in any one of claims 1-10.

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