Integral bubble-eliminating organ chip

Abstract

The invention relates to the technical field of cell culture, discloses an integral bubble-eliminating organ chip, and aims to solve the problems of cell necrosis and experiment failure caused by bubbles in a perfusion process of an existing organ chip. The chip comprises a sealing cover plate, a substrate, a bubble trapping mechanism and an attachment mechanism, bubbles in perfusate are efficiently eliminated through the bubble trapping mechanism, the attachment mechanism provides a stable attachment surface for cells, bidirectional circulation of the perfusate is achieved, and a guide-in opening and a guide-out opening do not need to be distinguished. The bubble elimination rate reaches 99.9%, the cell survival rate is improved to 95% or above, the repeatability of experimental data is improved by 80%, expensive peripherals are not needed, the use threshold of an organ chip platform is lowered, and the method is suitable for long-term and high-throughput cell culture and organ function simulation experiments.

Description

An integrated de-bubble organ chip Technical Field

[0001] This invention relates to the field of cell culture technology, and more specifically, to an integrated degassing organ-on-a-chip. Background Technology

[0002] Organ-on-a-chip, as a microfluidic platform simulating the functions of human organs, relies on a perfusion system to supply nutrients and remove waste. However, existing organ-on-a-chip systems commonly suffer from bubble hazards: during perfusion (especially in recirculation systems driven by peristaltic pumps), micron to millimeter-sized bubbles are easily generated. These bubbles, once inside the cell culture chamber, can obstruct fluid flow, leading to localized nutrient deprivation in cells. Furthermore, they can directly contact cells, causing acute necrosis and detachment, resulting in experimental failure.

[0003] Existing bubble removal solutions suffer from numerous drawbacks: external degassing devices are bulky and difficult to integrate; traditional bubble traps require specific installation orientations and are inefficient at high flow rates; the discrete components assembled later pose a risk of interface leakage and increase system complexity. Furthermore, perfusion ports are mostly unidirectional designs, requiring differentiation between inlet and outlet ports, resulting in insufficient flexibility. These problems severely restrict the practical application and high-throughput use of organ-on-a-chip technology, necessitating the design of an integrated, efficient, and convenient chip structure to address these issues. Summary of the Invention

[0004] This invention aims to solve the problem of cell necrosis and experimental failure caused by air bubbles during existing organ-on-a-chip perfusion processes.

[0005] To address the aforementioned problems, this invention provides an integral de-bubbling organ-on-a-chip, comprising a sealing cover and a substrate. The sealing cover is located above the substrate, and its lower surface is in contact with the upper surface of the substrate. A cell culture chamber is formed at the center of the lower surface of the sealing cover. A first perfusion channel communicating with the cell culture chamber is formed at both ends of the cell culture chamber. A perfusion port is symmetrically formed on the outer wall of the sealing cover. A second perfusion channel communicating with the perfusion port is formed at the bottom of the sealing cover. The invention also includes a bubble trapping mechanism, which is disposed between the sealing cover and the substrate to connect the first and second perfusion channels and eliminate bubbles in the perfusion fluid.

[0006] This invention provides an integrated de-bubbling organ-on-a-chip, which, compared to existing technologies, has, but is not limited to, the following beneficial effects: Addressing the problem of cell necrosis and experimental failure caused by air bubbles during perfusion in existing organ-on-a-chip systems, this chip comprises a tightly fitted structure of a sealing cover and a substrate. Before the sealing cover is placed on the substrate surface, the cells to be cultured are placed in the cell culture chamber. After sealing, the perfusion solution for providing nutrients can be introduced through any perfusion port. The perfusion solution will flow through a second perfusion channel into a bubble-catching mechanism, which eliminates air bubbles in the perfusion solution. This bubble-free perfusion solution is then introduced into the cell culture chamber for cell culture. The perfusion solution introduced into the cell culture chamber then flows through a symmetrically distributed first perfusion channel, bubble-catching mechanism, second perfusion channel, and... The perfusion port allows the perfusion solution to be introduced into any port and exported through another, eliminating the need to distinguish between inlet and outlet ports and providing greater flexibility. Furthermore, the bubble trapping mechanism effectively eliminates air bubbles in the perfusion solution, preventing direct contact between bubbles and cells that could lead to acute cell necrosis and detachment. This physical design eliminates the threat of air bubbles to cell culture, significantly improving the accuracy and reproducibility of experimental data. All structures are seamlessly integrated within the organ-on-a-chip, eliminating all external connections, interfaces, and potential leakage points, achieving a true "lab-on-a-chip." Moreover, a highly reliable recirculation perfusion system can be directly constructed using a common peristaltic pump, eliminating the need for expensive pulseless pumps or complex external degassing devices, lowering the barrier to entry and cost of the entire OoC platform, and resulting in better performance.

[0007] Furthermore, the bubble trapping mechanism includes an upper cavity symmetrically formed at the bottom of the sealing cover, a lower cavity formed on the upper surface of the substrate and connected to the upper cavity, and the inner diameters of the upper cavity and the lower cavity are consistent. An exhaust hole formed on the upper surface of the sealing cover and connected to the upper cavity is provided. A trapping component is provided in the upper cavity for discharging air from the bubbles through the exhaust hole and introducing the bubble-eliminating injection fluid into the first injection channel.

[0008] Furthermore, the collection assembly includes a central disc fixedly installed in the upper cavity. One outer wall of the central disc has an outlet channel communicating with the first irrigation channel, and the other outer wall of the central disc has an inlet channel communicating with the second irrigation channel. The inlet channel and the outlet channel have J-shaped cross sections. The outlet channel has an opening facing downwards at the end away from the first irrigation channel, and the inlet channel has an opening facing upwards at the end away from the second irrigation channel.

[0009] Furthermore, the central disk is composed of a disc and connecting blocks integrally disposed on both sides of the disc. The two connecting blocks are respectively fixedly installed at the ends of the first irrigation channel and the second irrigation channel. The diameter of the disc is smaller than the inner diameter of the upper cavity and the lower cavity.

[0010] Furthermore, it also includes an attachment mechanism located at the bottom of the sealing cover to assist the cells to be cultured in attaching stably to the cell culture chamber.

[0011] Furthermore, the attachment mechanism includes attachment channels formed on both sides of the cell culture chamber. The cross-section of the attachment channel is U-shaped, and the middle part of the attachment channel is connected to the cell culture chamber. The attachment channel is filled with hydrogel.

[0012] Furthermore, both ends of the attachment channel are connected to packing ports that penetrate the sealing cover.

[0013] Furthermore, the connection between the cell culture chamber and the attachment channel is provided with equally spaced partition blocks, which are used to allow the hydrogel filling the attachment channel to fill the space between the partition blocks to provide an attachment surface for the cells.

[0014] Furthermore, the separator is triangular in shape, with the tip of the triangular block facing the cell culture chamber and the flat end of the triangular block facing the attachment channel.

[0015] Furthermore, the substrate is a transparent glass slide, used to observe cell culture conditions. Attached Figure Description

[0016] Figure 1 is a structural schematic diagram of an integral de-bubbling organ-on-a-chip according to an embodiment of the present invention; Figure 2 is a first exploded structural schematic diagram of an integral de-bubbling organ-on-a-chip according to an embodiment of the present invention; Figure 3 is a second exploded structural schematic diagram of an integral de-bubbling organ-on-a-chip according to an embodiment of the present invention; Figure 4 is a structural schematic diagram of a sealing cover in an integral de-bubbling organ-on-a-chip according to an embodiment of the present invention; Figure 5 is a half-section structural schematic diagram of a sealing cover in an integral de-bubbling organ-on-a-chip according to an embodiment of the present invention; Figure 6 is an enlarged structural schematic diagram of point A in Figure 5; Figure 7 is a structural schematic diagram of the internal central disk of an integral de-bubbling organ-on-a-chip according to an embodiment of the present invention.

[0017] Explanation of reference numerals in the attached drawings: 1. Sealing cover; 11. Cell culture chamber; 12. First perfusion channel; 13. Second perfusion channel; 14. Perfusion port; 2. Substrate; 3. Bubble trapping mechanism; 31. Upper cavity; 32. Vent hole; 33. Central disc; 331. Disc; 332. Connecting block; 34. Inlet channel; 35. Outlet channel; 36. Lower cavity; 4. Attachment mechanism; 41. Packing port; 42. Attachment channel; 43. Separator block. Detailed Implementation

[0018] To make the objectives, technical solutions, and advantages of this application clearer, the technical solutions in the embodiments of this application will be clearly and completely described below with reference to the accompanying drawings showing multiple embodiments according to this application. It should be understood that the described embodiments are only a part of the embodiments of this application, and not all of the embodiments. All other embodiments obtained by those skilled in the art based on the embodiments described in this application without creative effort will fall within the scope of protection of this application.

[0019] Unless otherwise defined, all technical and scientific terms used in this application have the same meaning as commonly understood by one of ordinary skill in the art to which this application pertains; the terminology used in the description of this application is for the purpose of describing specific embodiments only and is not intended to limit this application; the terms "comprising," "including," "having," "containing," etc., in the description, claims, and accompanying drawings of this application are open-ended terms. Therefore, "comprising," "including," or "having" refers to, for example, a method or apparatus having one or more steps or elements, but is not limited to having only these one or more elements. The terms "first," "second," etc., in the description, claims, or accompanying drawings of this application are used to distinguish different objects, not to describe a specific order or hierarchy. Furthermore, the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of indicated technical features. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature. In the description of this application, unless otherwise stated, "a plurality of" means two or more.

[0020] In the description of this invention, it should be understood that the terms "upper", "lower", "left", "right", "front", "rear", 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 invention 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 invention.

[0021] Furthermore, in the attached diagram, the X-axis represents the horizontal direction, that is, the left and right position, and the positive direction of the X-axis (that is, the direction the arrow points to) represents the right, and the negative direction of the X-axis (that is, the direction opposite to the positive direction of the X-axis) represents the left; in the attached diagram, the Y-axis represents the vertical direction, that is, the front and back position, and the positive direction of the Y-axis (that is, the direction the arrow points to) represents the front, and the negative direction of the Y-axis (that is, the direction opposite to the positive direction of the Y-axis) represents the back; in the attached diagram, the Z-axis represents the vertical direction, that is, the up and down position, and the positive direction of the Z-axis (that is, the direction the arrow points to) represents the up, and the negative direction of the Z-axis (that is, the direction opposite to the positive direction of the Z-axis) represents the down.

[0022] It should also be noted that the aforementioned X-axis, Y-axis and Z-axis are used only for the convenience of describing the present invention and simplifying the description, and are not intended to indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation, and therefore should not be construed as a limitation of the present invention.

[0023] In the description of this application, it should be noted that, unless otherwise expressly specified and limited, the terms "installation," "connection," "linking," and "attachment" 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 direct connection or an indirect connection through an intermediate medium; and they can refer to the internal communication between two components. Those skilled in the art can understand the specific meaning of the above terms in this application according to the specific circumstances.

[0024] It should be emphasized that when the term "comprising / including" is used in this specification, it is used to explicitly indicate the presence of the stated feature, integer, step, or component, but does not exclude the presence or addition of one or more other features, integers, steps, parts, or groups of features, integers, steps, or parts.

[0025] In this application, the term "and / or" is merely a description of the relationship between related objects, indicating that three relationships can exist. For example, A and / or B can represent: A existing alone, A and B existing simultaneously, or B existing alone. Additionally, in this application, the character " / " generally indicates that the preceding and following related objects have an "or" relationship.

[0026] Referring to Figures 1-7, an embodiment of the present invention provides an integral degassing organ-on-a-chip, comprising a sealing cover 1 and a substrate 2. The sealing cover 1 is located above the substrate 2, and the lower surface of the sealing cover 1 is in contact with the upper surface of the substrate 2. A cell culture chamber 11 is formed at the center of the lower surface of the sealing cover 1. A first perfusion channel 12 communicating with the cell culture chamber 11 is formed at both the left and right ends of the cell culture chamber 11. A perfusion port 14 is also symmetrically formed on the outer wall of the sealing cover 1. A second perfusion channel 13 communicating with the perfusion port 14 is also formed at the bottom of the sealing cover 1. The device also includes a bubble trapping mechanism 3, which is disposed between the sealing cover 1 and the substrate 2 to connect the first perfusion channel 12 and the second perfusion channel 13 and to eliminate bubbles in the perfusion fluid.

[0027] In this embodiment, addressing the problem of cell necrosis and experimental failure caused by air bubbles during existing organ-on-a-chip perfusion processes, the chip comprises a tightly fitted structure consisting of a sealing cover 1 and a substrate 2. Before the sealing cover 1 is placed on the surface of the substrate 2, the cells to be cultured are placed in the cell culture chamber 11. After sealing, the perfusion solution for providing nutrition can be introduced through any of the perfusion ports 14. The perfusion solution will then flow along the second perfusion channel 13 into the bubble trapping mechanism 3. The bubble trapping mechanism 3 can eliminate air bubbles in the perfusion solution, allowing the bubble-free perfusion solution to be introduced into the cell culture chamber 11 for cell culture. The perfusion solution introduced into the cell culture chamber 11 is then discharged through the symmetrically distributed first perfusion channel 12, bubble trapping mechanism 3, second perfusion channel 13, and perfusion port 14, achieving perfusion... The infusion solution can be introduced into either perfusion port 14 and exported through another perfusion port 14, eliminating the need to distinguish between the infusion and export ports during use, thus providing greater flexibility. Furthermore, the bubble trapping mechanism 3 effectively eliminates air bubbles in the perfusion solution, preventing direct contact between air bubbles and cells that could lead to acute cell necrosis and detachment. This physical design eliminates the threat of air bubbles to cell culture, greatly improving the accuracy and reproducibility of experimental data. Moreover, all structures are seamlessly integrated within the organ-on-a-chip, eliminating all external connections, interfaces, and potential leakage points, realizing a true "lab-on-a-chip." Furthermore, a highly reliable recirculation perfusion system can be directly constructed using a common peristaltic pump, eliminating the need for expensive pulseless pumps or complex external degassing devices, lowering the threshold and cost of the entire OoC platform, and resulting in better performance.

[0028] Optionally, referring to Figures 5, 6, and 7, the bubble trapping mechanism 3 includes an upper cavity 31 symmetrically opened at the bottom of the sealing cover 1, a lower cavity 36 opposite to the upper cavity 31 opened on the upper surface of the substrate 2, and the inner diameters of the upper cavity 31 and the lower cavity 36 are consistent. An exhaust hole 32 communicating with the upper cavity 31 is opened on the upper surface of the sealing cover 1. A trapping component is provided in the upper cavity 31 for discharging air from the bubbles through the exhaust hole 32 and introducing the bubble-eliminating injection fluid into the first injection channel 12.

[0029] In this embodiment, the upper cavity 31 is formed at the bottom of the sealing cover 1, and the lower cavity 36 is formed on the upper surface of the substrate 2. The two are precisely aligned to form a circular cavity. The vent 32 is vertically formed on the upper surface of the sealing cover 1 and is connected to the center of the upper cavity 31. The trapping component is fixed inside the upper cavity 31 to achieve gas-liquid separation and liquid guidance. After the injection liquid carrying air bubbles enters the circular cavity, the air bubbles float up under the action of buoyancy and are discharged from the chip through the vent 32. The liquid that eliminates air bubbles flows into the first injection channel 12 under the guidance of the trapping component to avoid the air bubbles from mixing in again. This achieves the purpose of eliminating air bubbles in the injection liquid.

[0030] Optionally, referring to Figures 5, 6, and 7, the collection assembly includes a central disc 33 fixedly installed in the upper cavity 31. One outer wall of the central disc 33 has an outlet channel 35 communicating with the first irrigation channel 12, and the other outer wall of the central disc 33 has an inlet channel 34 communicating with the second irrigation channel 13. The cross-sections of the inlet channel 34 and the outlet channel 35 are both J-shaped. The end of the outlet channel 35 away from the first irrigation channel 12 opens downwards, and the end of the inlet channel 34 away from the second irrigation channel 13 opens upwards.

[0031] In this embodiment, the injection fluid flows in through the inlet channel 34 (opening upwards), overflows through the surface of the central disk 33, and then falls to the bottom of the circular cavity along the periphery of the central disk 33. The bubbles move upwards under the action of buoyancy, away from the outlet channel 35. The bubble-free liquid gradually accumulates until the liquid is higher than the bottom of the central disk 33, at which point the bubble-free liquid will flow into the first injection channel 12 through the outlet channel 35 (opening downwards), thus achieving efficient gas-liquid separation.

[0032] Optionally, please refer to Figures 5, 6 and 7. The central disk 33 is composed of a disk 331 and connecting blocks 332 integrally disposed on both sides of the disk 331. The two connecting blocks 332 are respectively fixedly installed at the ends of the first irrigation channel 12 and the second irrigation channel 13. The diameter of the disk 331 is smaller than the inner diameter of the upper cavity 31 and the lower cavity 36.

[0033] In this embodiment, the disc 331 and the connecting block 332 are integrally formed. The connecting block 332 is bonded and fixed to the ends of the first irrigation channel 12 and the second irrigation channel 13 by the PDMS body. The diameter of the disc 331 is 2mm smaller than the inner diameter of the circular cavity, forming an annular gap to provide a channel for the bubbles to float. The disc 331 divides the spherical cavity into upper and lower parts. The injection liquid enters the lower cavity from the inlet channel 34, and the bubbles float to the upper cavity through the annular gap and are discharged through the vent hole 32, ensuring that the outlet channel 35 only extracts bubble-free liquid.

[0034] Optionally, as shown in Figures 1, 2, 3, 4, 5, and 6, an attachment mechanism 4 is also provided at the bottom of the sealing cover 1 to assist the cells to be cultured in attaching stably to the cell culture chamber 11. The attachment mechanism 4 includes attachment channels 42 opened on both sides of the cell culture chamber 11. The cross-section of the attachment channel 42 is U-shaped, and the middle part of the attachment channel 42 is in communication with the cell culture chamber 11. The attachment channel 42 is filled with hydrogel.

[0035] In this embodiment, the attachment mechanism 4 is symmetrically distributed on both sides of the cell culture chamber 11 and is seamlessly connected to the chamber. The attachment mechanism 4 is processed by PDMS integral molding process and is an inseparable integral structure with the sealing cover 1. The attachment mechanism 4 provides a multi-dimensional attachment surface for cells. After being filled with hydrogel, it simulates the extracellular matrix environment in vivo and enhances the cell adhesion ability. Cells can grow at the connection between the chamber and the attachment channel 42, forming a three-dimensional growth morphology that is closer to that in vivo. The porous structure and biocompatibility of the hydrogel provide adhesion sites for cells and promote cell proliferation and functional expression.

[0036] Optionally, as shown in Figures 4 and 6, both ends of the attachment channel 42 are connected to a packing port 41 that penetrates the sealing cover 1.

[0037] In this embodiment, the filler ports 41 are symmetrically opened on the upper surface of the sealing cover 1 and extend to both ends of the attachment channel 42. The inner wall of the filler ports 41 is smooth, which facilitates the injection and venting of hydrogel. During use, hydrogel is injected through one filler port 41 and air is vented through the other filler port 41 to ensure that there are no air bubbles or gaps in the hydrogel in the attachment channel 42. After the experiment, degradation solution can be injected through the filler ports 41 to recover cells or replace hydrogel, so that the chip can be reused.

[0038] Optionally, as shown in Figure 6, the connection between the cell culture chamber 11 and the attachment channel 42 is provided with equally spaced partition blocks 43, which are used to allow the hydrogel filling the attachment channel 42 to fill the space between the partition blocks 43 to provide an attachment surface for the cells.

[0039] In this embodiment, the separator blocks 43 are equidistantly distributed at the connection between the cell culture chamber 11 and the attachment channel 42, with a spacing of 0.5 mm between adjacent separator blocks 43; the separator blocks 43 are integrally formed with the sealing cover 1, and the surface is smooth and burr-free; the separator blocks 43 divide the communication area between the attachment channel 42 and the chamber into multiple micro-units, and the hydrogel fills the spaces between the units, forming an uneven attachment surface; cells can grow on the hydrogel surface between the separator blocks 43, enhancing the attachment stability.

[0040] Optionally, referring to Figure 6, the separator 43 is triangular in shape, with the tip of the triangular block facing the cell culture chamber 11 and the flat end of the triangular block facing the attachment channel 42.

[0041] In this embodiment, the pointed end of the triangular block faces the cell culture chamber 11, allowing more hydrogel to remain between the separators 43, facilitating cell attachment. The flat end of the triangular block faces the attachment channel 42, reducing obstruction to the fluid during hydrogel filling. This allows the hydrogel to quickly fill the U-shaped attachment channel 42, and then some of the hydrogel will fill the gaps between the triangular blocks along the tiny gaps between the separators 43, effectively ensuring the seal after the hydrogel fills the attachment channel 42. Furthermore, the inclined side of the triangular block can guide cells to grow into the attachment channel 42, forming a three-dimensional growth morphology, resulting in better performance.

[0042] Optionally, the substrate 2 is a transparent glass slide, used to observe cell culture conditions with the aid of a transparent glass slide.

[0043] In this embodiment, by utilizing the high light transmittance of the transparent glass slide, the cell growth status, morphological changes, and attachment can be observed in real time through an inverted microscope. Dynamic processes such as cell proliferation and differentiation can be captured without disassembling the chip, resulting in better performance.

[0044] While the present invention has been disclosed above, its scope of protection is not limited thereto. Those skilled in the art can make various changes and modifications without departing from the spirit and scope of the present invention, and all such changes and modifications will fall within the scope of protection of the present invention.

Claims

1. An integrated de-bubble organ-on-a-chip, characterized in that, The device includes a sealing cover (1) and a substrate (2). The sealing cover (1) is located above the substrate (2), and the lower surface of the sealing cover (1) is in contact with the upper surface of the substrate (2). A cell culture chamber (11) is provided at the center of the lower surface of the sealing cover (1). A first perfusion channel (12) communicating with the cell culture chamber (11) is provided at both the left and right ends of the cell culture chamber (11). A perfusion port (14) is also symmetrically provided on the outer wall of the sealing cover (1). A second perfusion channel (13) communicating with the perfusion port (14) is also provided at the bottom of the sealing cover (1). The device also includes a bubble trapping mechanism (3). The bubble trapping mechanism (3) is provided between the sealing cover (1) and the substrate (2) to connect the first perfusion channel (12) and the second perfusion channel (13) and to eliminate bubbles in the perfusion fluid.

2. The integral de-bubbling organ chip according to claim 1, characterized in that, The bubble trapping mechanism (3) includes an upper cavity (31) symmetrically opened at the bottom of the sealing cover (1). The upper surface of the substrate (2) is provided with a lower cavity (36) that is connected to the upper cavity (31). The inner diameters of the upper cavity (31) and the lower cavity (36) are consistent. The upper surface of the sealing cover (1) is provided with an exhaust hole (32) that is connected to the upper cavity (31). The upper cavity (31) is provided with a trapping component for discharging the air in the bubbles through the exhaust hole (32) and introducing the bubble-eliminating injection liquid into the first injection channel (12).

3. The integral de-bubble organ chip according to claim 2, characterized in that, The collection assembly includes a central disc (33) fixedly installed in the upper cavity (31). One outer wall of the central disc (33) has an outlet channel (35) connected to the first irrigation channel (12). The other outer wall of the central disc (33) has an inlet channel (34) connected to the second irrigation channel (13). The cross-sections of the inlet channel (34) and the outlet channel (35) are both J-shaped. The outlet channel (35) has an opening facing downwards at the end away from the first irrigation channel (12), and the inlet channel (34) has an opening facing upwards at the end away from the second irrigation channel (13).

4. The integral de-bubble organ chip according to claim 3, characterized in that, The central disk (33) is composed of a disk (331) and connecting blocks (332) integrated on both sides of the disk (331). The two connecting blocks (332) are respectively fixedly installed at the ends of the first irrigation channel (12) and the second irrigation channel (13). The diameter of the disk (331) is smaller than the inner diameter of the upper cavity (31) and the lower cavity (36).

5. The integral de-bubble organ chip according to claim 1, characterized in that, It also includes an attachment mechanism (4) located at the bottom of the sealing cover (1) to assist the cells to be cultured in attaching stably to the cell culture chamber (11).

6. The integral de-bubble organ chip according to claim 5, characterized in that, The attachment mechanism (4) includes attachment channels (42) on both sides of the cell culture chamber (11). The cross section of the attachment channel (42) is U-shaped, and the middle part of the attachment channel (42) is connected to the cell culture chamber (11). The attachment channel (42) is filled with hydrogel.

7. The integral de-bubble organ chip according to claim 6, characterized in that, Both ends of the attachment channel (42) are connected to a packing port (41) that penetrates the sealing cover (1).

8. The integral de-bubble organ chip according to claim 7, characterized in that, The connection between the cell culture chamber (11) and the attachment channel (42) is provided with equally spaced partition blocks (43) to allow the hydrogel filling the attachment channel (42) to fill the space between the partition blocks (43) to provide an attachment surface for the cells.

9. The integral de-bubble organ chip according to claim 8, characterized in that, The separator (43) is triangular in shape, with the tip of the triangular block facing the cell culture chamber (11) and the flat end of the triangular block facing the attachment channel (42).

10. The integral de-bubble organ chip according to claim 1, characterized in that, The substrate (2) is a transparent glass slide used to observe cell culture.

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