Microfluidic chip

Abstract

The application discloses a microfluidic chip, which comprises a main body, a liquid inlet storage structure, a liquid outlet storage structure and a liquid blocking structure. The main body is provided with hydrogel channels and culture fluid channels which are spaced apart and partially communicated; the liquid inlet storage structure is provided with a glue storage pool and a liquid inlet storage pool, the glue storage pool is communicated with one end of the hydrogel channels, and the liquid inlet storage pool is communicated with one end of the culture fluid channels; the liquid outlet storage structure is provided with a glue outlet storage pool and a liquid outlet storage pool, the glue outlet storage pool is communicated with the other end of the hydrogel channels, and the liquid outlet storage pool is communicated with the other end of the culture fluid channels; the liquid blocking structure is arranged in the main body and located between the hydrogel channels and the culture fluid channels, and a communication port is formed between the liquid blocking structure and the main body. The above-mentioned embodiments of the application can ensure the continuity and stability of the liquid entering the corresponding channels under the cooperation of the liquid inlet storage structure and the liquid outlet storage structure, and can prevent the discharged liquid from overflowing, and reduce the possibility of splashing or turbulent flow of the discharged liquid.

Description

[0001] This application claims priority to Chinese Patent Application No. 2022109108923, filed on July 29, 2022, entitled “Microfluidic Chip”, the entire contents of which are incorporated herein by reference. Technical Field

[0002] This application relates to the field of biological cell culture technology, and in particular to a microfluidic chip. Background Technology

[0003] In related technological fields, microfluidic chips can be used for the cultivation of three-dimensional cells, tissues, or organs. Utilizing microfluidic technology, various cell types are cultured on molds composed of specific materials to simulate the physiological and pathological processes of tissues and organs. By constructing the smallest functional units at the tissue and organ level, multicellular structures, tissue-tissue interfaces, physicochemical properties, mechanical cues, and vascular structures can be summarized. As a novel in vitro model, microfluidic chips have better application prospects compared to traditional in vitro models.

[0004] However, the fluid continuity and stability within traditional microfluidic chips are poor, requiring external devices for assistance. Without the intervention of external devices, the fluid between the injection and discharge ends cannot flow stably. Summary of the Invention

[0005] This application provides a microfluidic chip that, with the cooperation of an inlet liquid storage structure and an outlet liquid storage structure, ensures the continuity and stability of liquid entering the corresponding channel, and prevents the discharged liquid from overflowing, reducing the possibility of splashing or turbulence.

[0006] In a first aspect, embodiments of this application provide a microfluidic chip, comprising a main body, a liquid inlet reservoir, a liquid outlet reservoir, and a liquid blocking structure. The main body has mutually spaced and partially connected hydrogel channels and a culture fluid channel; the liquid inlet reservoir has a hydrogel inlet reservoir and a liquid inlet reservoir, the hydrogel inlet reservoir being connected to one end of the hydrogel channel and the liquid outlet reservoir being connected to one end of the culture fluid channel; the liquid outlet reservoir has a liquid outlet reservoir and a liquid outlet reservoir, the liquid outlet reservoir being connected to the other end of the hydrogel channel and the liquid outlet reservoir being connected to the other end of the culture fluid channel; the liquid blocking structure is disposed within the main body and located between the hydrogel channels and the culture fluid channel, and a communication port is formed between the liquid blocking structure and the main body;

[0007] The hydrogel can enter the hydrogel channel through the inlet reservoir and exit from the outlet reservoir. The hydrogel in the hydrogel channel can be blocked by the liquid-blocking structure and cannot flow into the culture fluid channel 112. The culture fluid can enter the culture fluid channel through the inlet reservoir and exit from the outlet reservoir. The culture fluid can exchange substances with the hydrogel through the connecting port.

[0008] The liquid inlet storage structure has a cylindrical liquid inlet storage space, which is divided into an inlet storage tank and the inlet storage tank of equal capacity; the liquid outlet storage structure has a cylindrical liquid outlet storage space, which is divided into an outlet storage tank and the outlet storage tank of equal capacity.

[0009] In some embodiments, the number of both the hydrogel channel and the culture fluid channel is one; the liquid inlet storage structure has a liquid inlet storage space, the liquid inlet storage structure includes a first liquid inlet partition plate, the first liquid inlet partition plate divides the liquid inlet storage space into a hydrogel inlet storage tank and a liquid inlet storage tank of equal capacity; the liquid outlet storage structure has a liquid outlet storage space, the liquid outlet storage structure further includes a first liquid outlet partition plate, the first liquid outlet partition plate divides the liquid outlet storage space into a hydrogel outlet storage tank and a liquid outlet storage tank of equal capacity.

[0010] In some embodiments, the number of hydrogel channels is one, the number of culture fluid channels is two, the hydrogel channel is located between the two culture fluid channels, and the liquid-blocking structure is provided between each culture fluid channel and the hydrogel channel;

[0011] The liquid inlet storage structure has a cylindrical liquid inlet storage space. The liquid inlet storage structure includes three second liquid inlet partition plates. The three second liquid inlet partition plates are disposed in the liquid inlet storage space. One end of the three second liquid inlet partition plates is connected, and the other end is connected to the inner wall of the liquid inlet storage structure in a divergent manner. The included angle between any two adjacent second liquid inlet partition plates is 120 degrees, so as to divide the cylindrical liquid inlet storage space into one glue inlet storage tank and two liquid inlet storage tanks with equal capacity.

[0012] The liquid outlet storage structure has a cylindrical liquid outlet storage space. The liquid outlet storage structure includes three second liquid outlet partition plates, which are disposed in the liquid outlet storage space. One end of the three second liquid outlet partition plates is connected, and the other end is connected to the inner wall of the liquid outlet storage structure in a divergent manner. The included angle between any two adjacent second liquid outlet partition plates is 120 degrees, so as to divide the cylindrical liquid outlet storage space into one liquid outlet storage pool and two liquid outlet storage pools of equal capacity.

[0013] In some embodiments, the length of the hydrogel channel is greater than the length of any one of the culture fluid channels, the two culture fluid channels are of equal length, and both ends of the hydrogel channel protrude relative to the culture fluid channels; the inlet reservoir is further away from the outlet reservoir structure than the two inlet reservoirs, so that the inlet reservoir is connected to the protruding end of the hydrogel channel relative to the culture fluid channel; the outlet reservoir is further away from the inlet reservoir structure than the two outlet reservoirs, so that the outlet reservoir is connected to the protruding end of the hydrogel channel relative to the culture fluid channel.

[0014] In some embodiments, the bottom of the glue inlet reservoir has a glue inlet, the flow cross-section of which is smaller than the flow cross-section of the glue inlet reservoir; the bottom of the liquid inlet reservoir has a liquid inlet, the flow cross-section of which is smaller than the flow cross-section of the liquid inlet reservoir; the bottom of the glue outlet reservoir has a glue outlet, the flow cross-section of which is smaller than the flow cross-section of the glue outlet reservoir; and the bottom of the liquid outlet reservoir has a liquid outlet, the flow cross-section of which is smaller than the flow cross-section of the liquid outlet reservoir.

[0015] In some embodiments, the main body includes a first substrate, a second substrate, and a third substrate; the first substrate has the hydrogel channel and the culture fluid channel; the second substrate has the gel inlet, the liquid inlet, the gel outlet, and the liquid outlet; the third substrate has the liquid inlet storage structure and the liquid outlet storage structure; wherein the liquid blocking structure is disposed on the first substrate and / or the second substrate.

[0016] In some embodiments, the side of the second substrate facing the first substrate is covered with a hydrophilic film or subjected to a hydrophilic surface treatment, or the entire second substrate is subjected to a hydrophilic surface treatment.

[0017] In some embodiments, the liquid-blocking structure includes a first liquid-blocking element and a second liquid-blocking element spaced apart, the hydrogel channel and the culture fluid channel are separated by the first liquid-blocking element and the second liquid-blocking element, the first liquid-blocking element is disposed on the surface of the first substrate facing the second substrate, the second liquid-blocking element is disposed on the surface of the second substrate facing the first substrate, and the communication port is formed between the first liquid-blocking element and the second liquid-blocking element.

[0018] In some embodiments, the first liquid-blocking element is at least one of a groove, a baffle, a boss, a hydrophobic film, and a grid; the second liquid-blocking element is at least one of a groove, a baffle, a boss, a hydrophobic film, and a grid.

[0019] In some embodiments, the width of the hydrogel channel is a, 200 μm ≤ a ≤ 600 μm; the width of the culture fluid channel is b, 300 μm ≤ b ≤ 700 μm; and the height of the hydrogel channel and the culture fluid channel is h, 200 μm ≤ h ≤ 400 μm.

[0020] The microfluidic chip based on the embodiments of this application can ensure the continuity and stability of liquid entering the corresponding channel with the cooperation of the inlet liquid storage structure and the outlet liquid storage structure, and can prevent the outlet liquid from overflowing, reducing the possibility of splashing or turbulence of the outlet liquid. Attached Figure Description

[0021] To more clearly illustrate the technical solutions in the embodiments of this application or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0022] Figure 1 This is a schematic diagram of the structure of a microfluidic chip in one embodiment of this application, formed by combining three substrates.

[0023] Figure 2 This is a schematic diagram of the microfluidic chip integrally formed according to an embodiment of this application;

[0024] Figure 3 This is a top view of a microfluidic chip in one embodiment of the present application, which includes a hydrogel channel and two culture fluid channels.

[0025] Figure 4 This is a top view of a microfluidic chip in one embodiment of the present application, which includes a hydrogel channel and a culture fluid channel.

[0026] Figure 5This is a schematic diagram of the exploded structure of a microfluidic chip in one embodiment of this application, which includes a hydrogel channel and two culture fluid channels.

[0027] Figure 6 This is a schematic diagram of the structure of the first substrate in a microfluidic chip according to an embodiment of this application;

[0028] Figure 7 This is a schematic diagram of the structure of the second substrate near the first substrate in a microfluidic chip according to an embodiment of this application;

[0029] Figure 8 This is a schematic diagram of the structure of the third substrate in a microfluidic chip according to an embodiment of this application;

[0030] Figure 9 This is a cross-sectional view of a microfluidic chip in one embodiment of the present application, which includes a hydrogel channel and two culture fluid channels.

[0031] Figure 10 This is a schematic diagram of a microfluidic chip in one embodiment of the present application, showing a structure in which hydrogel and liquid-blocking structure cooperate to form a liquid-blocking boundary;

[0032] Figure 11 This is a schematic diagram of another structure in a microfluidic chip according to one embodiment of the present application, in which hydrogel and liquid-blocking structure cooperate to form a liquid-blocking boundary;

[0033] Figure label:

[0034] 100. Main body; 110. First substrate; 111. Hydrogel channel; 112. Culture fluid channel; 120. Second substrate; 121. Inlet; 122. Liquid inlet; 123. Outlet; 124. Liquid outlet; 130. Third substrate; 131. Liquid inlet storage structure; 1311. Inlet storage tank; 1312. Inlet storage tank; 1313. First inlet partition plate; 1314. Second inlet partition plate; 132. Outlet storage structure; 1321. Outlet storage tank; 1322. Outlet storage tank; 1323. First outlet partition plate; 1324. Second outlet partition plate; 140. Liquid blocking structure; 141. First liquid blocking component; 142. Second liquid blocking component; 143. Liquid blocking boundary. Detailed Implementation

[0035] To make the objectives, technical solutions, and advantages of this application clearer, the following detailed description is provided in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the scope of this application.

[0036] Reference Figures 1 to 11As shown, the first aspect of this application proposes a microfluidic chip, including a main body 100, a liquid inlet storage structure 131, a liquid outlet storage structure 132, and a liquid blocking structure 140. With the cooperation of the liquid inlet storage structure 131 and the liquid outlet storage structure 132, it can ensure the continuity and stability of liquid entering the corresponding channel, and can prevent the discharged liquid from overflowing, reducing the possibility of splashing or turbulence of the discharged liquid.

[0037] In some embodiments of this application, the body 100 of the microfluidic chip has mutually spaced and partially connected hydrogel channels 111 and culture fluid channels 112; the hydrogel channels 111 allow hydrogel to flow through, and the culture fluid channels 112 allow culture fluid for cell culture to flow through. The culture fluid can be gas, culture medium, or drug during cell culture. The mutually spaced and partially connected hydrogel channels 111 and culture fluid channels 112 allow the hydrogel channels 111 and culture fluid channels 112 to be independent of each other, while the liquid-blocking structure 140 described below allows the culture fluid in the culture fluid channel 112 to exchange substances with the hydrogel, or allows the culture fluid in two different culture fluid channels 112 to exchange substances through the hydrogel.

[0038] Reference Figures 1 to 4 The liquid inlet storage structure 131 can store a certain amount of liquid to be entered into the channel, ensuring the continuity of liquid entry into the corresponding channel. Specifically, the liquid inlet storage structure 131 may include a hydrogel inlet storage tank 1311 and a liquid inlet storage tank 1312. The hydrogel inlet storage tank 1311 can store liquids such as hydrogel and is correspondingly connected to the hydrogel channel 111. The liquid inlet storage tank 1312 can store liquids such as culture fluid and is correspondingly connected to the culture fluid channel 112. Specifically, the hydrogel inlet storage tank 1311 is connected to one end of the hydrogel channel 111, and the liquid inlet storage tank 1312 is connected to one end of the culture fluid channel 112.

[0039] Reference Figures 1 to 4 The liquid outlet storage structure 132 can store a certain amount of liquid to be discharged from the channel, ensuring that the discharged liquid will not overflow and reducing the possibility of splashing or turbulence. Specifically, the liquid outlet storage structure 132 can include a gel dispensing reservoir 1321 and a liquid outlet storage tank 1322. The gel dispensing reservoir 1321 can also store liquids such as hydrogel and is connected to the hydrogel channel 111. The liquid outlet storage tank 1322 can also store liquids such as culture fluid and is connected to the culture fluid channel 112. Specifically, the gel dispensing reservoir 1321 is connected to the other end of the hydrogel channel 111, and the liquid outlet storage tank 1322 is connected to the other end of the culture fluid channel 112.

[0040] Reference Figures 5 to 7The liquid-blocking structure 140 can confine the hydrogel within the hydrogel channel 111 to the hydrogel channel 111, reducing or even preventing the possibility of hydrogel flowing into the culture fluid channel 112. Specifically, the liquid-blocking structure 140 is disposed within the main body 100 and located between the hydrogel channel 111 and the culture fluid channel 112, and a communication port 144 is formed between the liquid-blocking structure 140 and the main body 100.

[0041] It should be noted that the hydrogel can enter the hydrogel channel 111 through the inlet reservoir 1311 and can be discharged from the outlet reservoir 1321. The hydrogel in the hydrogel channel 111 can be blocked by the liquid-blocking structure 140 and cannot flow into the culture fluid channel 112. The culture fluid can enter the culture fluid channel 112 through the inlet reservoir 1312 and can be discharged from the outlet reservoir 1322. The culture fluid can exchange substances with the hydrogel through the connecting port 144.

[0042] The above-described embodiments of this application, with the cooperation of the inlet liquid storage structure 131 and the outlet liquid storage structure 132, can ensure the continuity and stability of liquid entering the corresponding channel, and can prevent the discharged liquid from overflowing, reducing the possibility of splashing or turbulence of the discharged liquid.

[0043] Reference Figure 4 In some embodiments of this application, the number of hydrogel channels 111 and culture fluid channels 112 is one each; the liquid inlet storage structure 131 has a liquid inlet storage space, and the liquid inlet storage structure 131 includes a first liquid inlet partition plate 1313, which divides the liquid inlet storage space into a hydrogel inlet storage pool 1311 and a liquid inlet storage pool 1312 of equal capacity; the liquid outlet storage structure 132 has a liquid outlet storage space, and the liquid outlet storage structure 132 also includes a first liquid outlet partition plate 1323, which divides the liquid outlet storage space into a hydrogel outlet storage pool 1321 and a liquid outlet storage pool 1322 of equal capacity.

[0044] Based on the above embodiments of this application, the number of hydrogel channels 111 and culture fluid channels 112 is one each. In this case, the liquid inlet storage structure 131 includes a hydrogel inlet storage tank 1311 and a liquid inlet storage tank 1312. The liquid inlet storage space of the liquid inlet storage structure 131 can be divided into two sub-spaces by a first liquid inlet partition plate 1313. One sub-space is the hydrogel inlet storage tank 1311, which is connected to one end of the hydrogel channel 111, and the other sub-space is the liquid inlet storage tank 1312, which is connected to one end of the culture fluid channel 112. The first liquid inlet partition plate 1313 can be positioned in the center to divide the liquid inlet storage space into the hydrogel inlet storage tank 1311 and the liquid inlet storage tank 1312, both of equal capacity.

[0045] It should be noted that, as needed, the first liquid inlet partition plate 1313 can also be eccentrically set to divide the liquid inlet storage space into a gel inlet storage pool 1311 and a liquid inlet storage pool 1312 with unequal capacities. Specifically, the capacity of the gel inlet storage pool 1311 can be smaller than that of the liquid inlet storage pool 1312. The hydrogel in the hydrogel channel 111 needs to be solidified during cell culture, so the liquid inlet volume can be controlled to be relatively small. Meanwhile, the culture fluid in the culture fluid channel 112 can be replenished with nutrients by flowing, so the liquid inlet volume is relatively large. This setting can make greater use of the usable space of the main body 100.

[0046] The liquid outlet storage structure 132 at this time includes a gel outlet storage tank 1321 and a liquid outlet storage tank 1322. The liquid outlet storage space of the liquid outlet storage structure 132 can be divided into two sub-spaces by a first liquid outlet partition plate 1323. One sub-space is the gel outlet storage tank 1321, which is connected to the other end of the hydrogel channel 111, and the other sub-space is the liquid outlet storage tank 1322, which is connected to the other end of the culture fluid channel 112. The first liquid outlet partition plate 1323 can be placed in the center to divide the liquid outlet storage space into the gel outlet storage tank 1321 and the liquid outlet storage tank 1322 with equal capacity.

[0047] It should be noted that, as needed, the first liquid outlet partition plate 1322 can also be eccentrically set to divide the liquid outlet storage space into a liquid outlet storage pool 1321 and a liquid outlet storage pool 1322 with unequal capacities. Specifically, the capacity of the liquid outlet storage pool 1321 can be smaller than that of the liquid outlet storage pool 1322. The hydrogel in the hydrogel channel 111 needs to be solidified during cell culture, so the liquid outlet volume can be controlled to be relatively small. Meanwhile, the culture fluid in the culture fluid channel 112 can be replenished with nutrients by flowing, resulting in a relatively large liquid outlet volume. This setting can make greater use of the usable space of the main body 100.

[0048] Reference Figures 1 to 3 , Figures 5 to 8 In some embodiments of this application, the number of hydrogel channels 111 is one, and the number of culture fluid channels 112 is two. The hydrogel channel 111 is located between the two culture fluid channels 112, and a liquid-blocking structure 140 is provided between each culture fluid channel 112 and the hydrogel channel 111.

[0049] The liquid inlet storage structure 131 has a cylindrical liquid inlet storage space. The liquid inlet storage structure 131 includes three second liquid inlet partition plates 1314. The three second liquid inlet partition plates 1314 are disposed in the liquid inlet storage space. One end of the three second liquid inlet partition plates 1314 is connected, and the other end is connected to the inner wall of the liquid inlet storage structure in a divergent manner. The included angle between any two adjacent second liquid inlet partition plates 1314 is 120 degrees, so as to divide the cylindrical liquid inlet storage space into one glue inlet storage pool 1311 and two liquid inlet storage pools 1312 with equal capacity.

[0050] The liquid outlet storage structure 132 has a cylindrical liquid outlet storage space. The liquid outlet storage structure 132 includes three second liquid outlet partition plates 1324. The three second liquid outlet partition plates 1324 are disposed in the liquid outlet storage space. One end of the three second liquid outlet partition plates 1324 is connected, and the other end is connected to the inner wall of the liquid outlet storage structure in a divergent manner. The included angle between any two adjacent second liquid outlet partition plates 1324 is 120 degrees, so as to divide the cylindrical liquid outlet storage space into one liquid outlet storage pool 1321 and two liquid outlet storage pools 1322 with equal capacity.

[0051] Based on the above embodiments of this application, one hydrogel channel 111 is provided, and two culture fluid channels 112 are provided. The hydrogel channel 111 is located between the two culture fluid channels 112, and a liquid-blocking structure 140 is provided between each culture fluid channel 112 and the hydrogel channel 111. In this way, after the hydrogel with the liquid-blocking structure 140 at the corresponding position is cured, the culture fluid in the two culture fluid channels 112 can exchange substances through the gaps or pores of the cured hydrogel.

[0052] Reference Figure 8 The liquid inlet storage structure 131 at this time includes one hydrogel inlet storage tank 1311 and two liquid inlet storage tanks 1312. The liquid inlet storage structure 131 has a cylindrical liquid inlet storage space, which can be divided into three sub-spaces by a second liquid inlet partition plate 1314. One sub-space is the hydrogel inlet storage tank 1311, which is connected to one end of the hydrogel channel 111; the other two sub-spaces are liquid inlet storage tanks 1312, which are connected to one end of the corresponding culture fluid channel 112. The second liquid inlet partition plate 1314 can be arranged in a ring with three equal parts, or the included angle between any two adjacent second liquid inlet partition plates 1314 is 120 degrees. One end of the three corresponding second liquid inlet partition plates 1314 is connected, and the other end is connected to the inner wall of the liquid inlet storage structure in a divergent manner, so as to divide the liquid inlet storage space into one hydrogel inlet storage tank 1311 and two liquid inlet storage tanks 1312 with equal capacity.

[0053] It should be noted that, as needed, the second liquid inlet partition plate 1314 can be set in a non-equal or parallel manner to divide the liquid inlet storage space into one liquid inlet storage pool 1311 and two liquid inlet storage pools 1312 with not necessarily equal capacities. Specifically, the capacity of the liquid inlet storage pool 1311 can be smaller than the capacity of the other two liquid inlet storage pools 1312. The hydrogel in the hydrogel channel 111 needs to be solidified during cell culture, so the liquid inlet volume can be controlled to be relatively small. The culture fluid in the culture fluid channel 112 can be replenished with nutrients by flowing, so the liquid inlet volume is relatively large. This arrangement can make greater use of the usable space of the main body 100.

[0054] The liquid outlet storage structure 132 at this time includes one gel outlet storage tank 1321 and two liquid outlet storage tanks 1322. The liquid outlet storage structure 132 has a cylindrical liquid outlet storage space, which can be divided into three sub-spaces by the second liquid outlet partition plate 1324. One sub-space is the gel outlet storage tank 1321, which is connected to the other end of the hydrogel channel 111; the other two sub-spaces are liquid outlet storage tanks 1322, which are connected to the other end of the corresponding culture fluid channel 112. The second liquid outlet partition plate 1324 can be arranged in a ring with three equal parts, or the included angle between any two adjacent second liquid outlet partition plates 1324 is 120 degrees. One end of the three corresponding second liquid outlet partition plates 1324 is connected, and the other end is connected to the inner wall of the liquid outlet storage structure in a divergent manner, so as to divide the liquid outlet storage space into one gel outlet storage tank 1321 and two liquid outlet storage tanks 1322 with equal capacity.

[0055] It should be noted that, as needed, the second liquid outlet partition plate 1324 can be set in a non-equal or parallel manner to divide the liquid outlet storage space into one liquid outlet storage pool 1321 and two liquid outlet storage pools 1322 with not necessarily equal capacities. Specifically, the capacity of the liquid outlet storage pool 1321 can be smaller than the capacity of the other two liquid outlet storage pools 1322. The hydrogel in the hydrogel channel 111 needs to be solidified during cell culture, so the liquid outlet volume can be controlled to be relatively small. Meanwhile, the culture fluid in the culture fluid channel 112 can be replenished with nutrients by flowing, resulting in a relatively large liquid outlet volume. This arrangement can make greater use of the usable space of the main body 100.

[0056] Reference Figure 5 In some embodiments of this application, the length of the hydrogel channel 111 is greater than the length of any culture fluid channel 112. This arrangement makes it easier to offset the ends of the hydrogel channel 111 from the ends of the culture fluid channel 112, thus making the ends of the two channels more easily distinguishable.

[0057] Specifically, the lengths of the two culture fluid channels 112 can be set to be equal, and the two ends of the hydrogel channel 111 can protrude relative to the culture fluid channel 112, so that the ends of the hydrogel channel 111 and the ends of the culture fluid channel 112 can be staggered. The above arrangement can make the inlet reservoir 1311 farther away from the outlet reservoir structure 132 than the two inlet reservoirs, so that the inlet reservoir 1311 is connected to the protruding end of the hydrogel channel 111 relative to the culture fluid channel 112. The outlet reservoir 1321 is farther away from the inlet reservoir structure 131 than the two outlet reservoirs, so that the outlet reservoir 1321 is connected to the protruding end of the hydrogel channel 111 relative to the culture fluid channel 112. This allows operators to easily distinguish between the glue inlet reservoir 1311 and the glue inlet reservoir 1312 in the liquid inlet reservoir structure 131, and also to easily distinguish between the glue outlet reservoir 1321 and the glue outlet reservoir 1322 in the liquid outlet reservoir structure 132.

[0058] It should be noted that the length mentioned above does not necessarily refer to the flow length of the fluid in the corresponding channel. The length mentioned above can be the relative length in the length direction of the microfluidic chip (when the top view is rectangular), specifically the straight-line distance between the two ends of the corresponding channel.

[0059] Reference Figure 5 In some embodiments of this application, the bottom of the glue inlet reservoir 1311 has a glue inlet 121, the flow cross-section of which is smaller than that of the glue inlet reservoir 1311; the bottom of the liquid inlet reservoir 1312 has a liquid inlet 122, the flow cross-section of which is smaller than that of the liquid inlet reservoir 1312; the bottom of the glue outlet reservoir 1321 has a glue outlet 123, the flow cross-section of which is smaller than that of the glue outlet reservoir 1321; and the bottom of the liquid outlet reservoir 1322 has a liquid outlet 124, the flow cross-section of which is smaller than that of the liquid outlet reservoir 1322.

[0060] Based on the above embodiments of this application, the inlet 121 allows the hydrogel in the hydrogel reservoir 1311 to flow into the hydrogel channel 111. When the flow cross-section of the inlet 121 is smaller than the flow cross-section of the hydrogel reservoir 1311, the speed at which the hydrogel flows into the hydrogel channel 111 can be slowed, improving the stability of the hydrogel flow. Similarly, the inlet 122 allows the culture fluid in the liquid reservoir 1312 to flow into the culture fluid channel 112. When the flow cross-section of the inlet 122 is smaller than the flow cross-section of the liquid reservoir 1312, the speed at which the culture fluid flows into the culture fluid channel 112 can be slowed, improving the stability of the culture fluid flow and ensuring a stable delivery of the culture fluid. The outlet 123 allows the hydrogel in the hydrogel channel 111 to be discharged into the outlet reservoir 1321. When the flow cross-section of the outlet 123 is smaller than that of the outlet reservoir 1321, the flow rate of the hydrogel towards the outlet reservoir 1321 can be slowed down, thus improving the stability of the hydrogel flow. The outlet 124 allows the culture fluid in the culture fluid channel 112 to flow into the outlet reservoir 1322. When the flow cross-section of the outlet 124 is smaller than that of the outlet reservoir 1322, the flow rate of the culture fluid towards the outlet reservoir 1322 can be slowed down, thus improving the stability of the culture fluid flow and ensuring that the culture fluid that has absorbed nutrients can be discharged stably.

[0061] Reference Figure 5 In some embodiments of this application, the main body 100 includes a first substrate 110, a second substrate 120, and a third substrate 130; the first substrate 110 has a hydrogel channel 111 and a culture fluid channel 112; the second substrate 120 has a gel inlet 121, a liquid inlet 122, a gel outlet 123, and a liquid outlet 124; the third substrate 130 has a liquid inlet storage structure 131 and a liquid outlet storage structure 132; wherein, a liquid blocking structure 140 is disposed on the first substrate 110 and / or the second substrate 120.

[0062] Based on the embodiments described above, the main body 100 can be a first substrate 110, a second substrate 120, and a third substrate 130 stacked and fixed as a whole. One end of the hydrogel channel 111, via its inlet 121, is connected to the liquid inlet storage structure 131, and the other end is connected to the liquid outlet storage structure 132 via its outlet 123. Similarly, one end of the culture fluid channel 112, via its inlet 122, is connected to the liquid inlet storage structure 131, and the other end is connected to the liquid outlet storage structure 132 via its outlet.

[0063] The liquid blocking structure 140 can be disposed on the first substrate 110 alone, or on the second substrate 120 alone, or the corresponding liquid blocking structure 140 can be disposed on both the first substrate 110 and the second substrate 120.

[0064] It should be noted that the first substrate 110, the second substrate 120 and the third substrate 130 are bonded and assembled into a whole by one of the following methods: hot pressing, ultrasonication, laser, and adhesive bonding.

[0065] The microfluidic chip is made of at least one of the following materials: plexiglass, polycarbonate, cyclic olefin copolymer, dimethyl methylphosphonate, or polystyrene.

[0066] Acrylic glass (Polymethyl methacrylate, abbreviated as PMMA). This transparent polymer material is chemically known as polymethyl methacrylate, a polymer compound formed by the polymerization of methyl methacrylate. It possesses characteristics such as a smooth surface, vibrant colors, low specific gravity, high strength, corrosion resistance, moisture resistance, sun resistance, good insulation properties, and good sound insulation. When acrylic glass is used in microfluidic chips, surface treatment is unnecessary, resulting in a lightweight, high-strength chip that is corrosion-resistant, moisture-resistant, sun-resistant, and exhibits stable performance.

[0067] Polycarbonate (PC) is a high molecular weight polymer containing carbonate groups in its molecular chain. Based on the structure of the ester groups, it can be classified into various types, including aliphatic, aromatic, and aliphatic-aromatic. Polycarbonate is a strong and tough thermoplastic resin. When used in microfluidic chips, it offers advantages such as high strength, high toughness, high temperature resistance, and relatively stable performance.

[0068] Cyclic olefin copolymers (COCs) are amorphous, transparent copolymers with cyclic olefin structures. Developed by TOPASADVANCED POLYMERS M.S. B.H., COCs are a high-quality, high-purity amorphous cyclic resin based on a proprietary metallocene catalyst technology. When used in microfluidic chips, COCs offer advantages such as lightweight construction and long lifespan.

[0069] Dimethyl methanephosphonate (COP) has strong mechanical properties, temperature resistance, and weather resistance. When polystyrene is used in microfluidic chips, it can have the characteristics of high strength and good heat resistance.

[0070] Polystyrene (PS) is a polymer synthesized from styrene monomers through a free radical polymerization reaction. Typical polystyrene is an amorphous random polymer with excellent thermal insulation, electrical insulation, and transparency, and a long-term operating temperature range of 0–70℃. When polystyrene is used in microfluidic chips, it can provide good thermal insulation, high transparency, and good electrical insulation.

[0071] It should be noted that the main body 100 of this application can also be formed by integral molding, specifically by additive manufacturing methods such as 3D printing.

[0072] In some embodiments of this application, the side of the second substrate 120 facing the first substrate 110 is covered with a hydrophilic film or subjected to a hydrophilic surface treatment, or the entire second substrate 120 is subjected to a hydrophilic surface treatment.

[0073] Based on the embodiments described above, the hydrophilic surface treatment can improve the fluid flow within the microfluidic chip. Specifically, a hydrophilic film can be applied to the side of the second substrate 120 facing the first substrate 110, or a hydrophilic surface treatment can be performed, or the entire second substrate 120 can be hydrophilically surface-treated. Alternatively, a hydrophilic film can be applied to the side of the first substrate 110 facing the second substrate 120, or a hydrophilic surface treatment can be performed as needed. Specifically, a portion of the inner surface of the hydrogel channel 111 and the culture fluid channel 112 can be hydrophilically surface-treated to improve the fluid flow within the microfluidic chip.

[0074] Reference Figures 5 to 11 In some embodiments of this application, the liquid-blocking structure 140 includes a first liquid-blocking element 141 and a second liquid-blocking element 142 spaced apart. The hydrogel channel 111 and the culture fluid channel 112 are separated by the first liquid-blocking element 141 and the second liquid-blocking element 142. The first liquid-blocking element 141 is disposed on the surface of the first substrate 110 facing the second substrate 120, and the second liquid-blocking element 142 is disposed on the surface of the second substrate 120 facing the first substrate 110. A communication port 144 is formed between the first liquid-blocking element 141 and the second liquid-blocking element 142.

[0075] Based on the above embodiments of this application, the cooperation of the first liquid blocking member 141 and the second liquid blocking member 142 can separate the hydrogel channel 111 and the culture fluid channel 112. Specifically, the first liquid blocking member 141 can be disposed on the surface of the first substrate 110 facing the second substrate 120, and the second liquid blocking member 142 can be disposed on the surface of the second substrate 120 facing the first substrate 110. The above-mentioned communication port 144 can facilitate the exchange of substances in the culture fluid in the culture fluid channel 112 after the hydrogel has solidified.

[0076] It should be noted that the first liquid-blocking element 141 forms a first liquid-blocking edge with the hydrogel, and the second liquid-blocking element 142 forms a second liquid-blocking edge with the hydrogel. A liquid-blocking boundary 143 can be formed between the first and second liquid-blocking edges. The liquid-blocking boundary 143 can be a curved liquid film. The liquid-blocking boundary 143 formed by the cooperation of the first and second liquid-blocking edges can confine the hydrogel within the hydrogel channel 111. When hydrogel is injected into the hydrogel channel 111, the hydrogel can flow from one end of the hydrogel channel 111 to the other end under the obstruction of the liquid-blocking boundary 143, and fill the communication position between the hydrogel channel 111 and the two culture fluid channels 112. During cell culture, the cultured cells in the two culture fluid channels 112 can exchange substances through the solidified hydrogel.

[0077] After the hydrogel between the two culture fluid channels 112 solidifies, it can form hydrogel strips that block the direct flow of culture fluid within the two culture fluid channels 112.

[0078] The microfluidic chip of this application can culture three-dimensional cells, tissues or organs, which may specifically include the heart, liver, lungs, spleen, kidneys, intestines, brain, nerves, blood vessels or skin, etc.

[0079] In some embodiments of this application, the first liquid-blocking element 141 is at least one of a groove, a baffle, a boss, a hydrophobic film, and a grid; the second liquid-blocking element 142 is at least one of a groove, a baffle, a boss, a hydrophobic film, and a grid. All of the above structures can generate a liquid-blocking effect using corresponding liquid-blocking elements, forming a corresponding liquid-blocking boundary 143. Corresponding structures can be set as needed.

[0080] Based on the above embodiments of this application, the first liquid blocking element 141 is at least one of a groove, a baffle, a boss, a hydrophobic film, and a grid; the second liquid blocking element 142 is at least one of a groove, a baffle, a boss, a hydrophobic film, and a grid.

[0081] In some embodiments of this application, the width of the hydrogel channel 111 is a, 200μm≤a≤600μm; the width of the culture fluid channel 112 is b, 300μm≤b≤700μm; and the height of the hydrogel channel 111 and the culture fluid channel 112 is h, 200μm≤h≤400μm.

[0082] Based on the above embodiments of this application, the above dimensions can ensure the formation of the liquid-blocking boundary 143 while allowing fluid to flow in the hydrogel channel 111 and the culture fluid channel 112, making the liquid-blocking boundary 143 less prone to damage.

[0083] Implementation Method 1

[0084] Taking a microfluidic chip as an example of an intestinal microchip, this scheme has two culture fluid channels: a first culture fluid channel and a second culture fluid channel. The first end of the first culture fluid channel is connected to a first inlet reservoir via a first inlet, and the second end is connected to a first outlet reservoir via a first outlet. Similarly, the first end of the second culture fluid channel is connected to a second inlet reservoir via a second inlet, and the second end is connected to a second outlet reservoir via a second outlet. Specific operating steps may include:

[0085] The second substrate is hydrophilically treated;

[0086] The first substrate, the second substrate, and the third substrate are stacked in sequence and assembled into a whole.

[0087] Before use, the chip (in this case, an intestinal chip) is sterilized.

[0088] When in use, Matrigel / collagen mixed hydrogel is added from the chip's gel reservoir and enters the channel hydrogel channel through the gel inlet. Due to the blocking effect of the liquid boundary, the hydrogel solution is confined to the hydrogel channel and does not flow into the other two culture fluid channels of the channel. Excess hydrogel flows out through the gel outlet and into the gel outlet reservoir.

[0089] After the hydrogel solidifies, it forms hydrogel strips.

[0090] Intestinal epithelial cells are added to the first inlet reservoir of the chip, entering and filling the first culture fluid channel through the first inlet port; the intestinal epithelial cells adhere to the hydrogel surface at the junction of the first culture fluid channel and the hydrogel channel; intestinal epithelial cell culture medium (culture fluid) is added to the first inlet reservoir of the chip, entering and filling the first culture fluid channel through the first inlet port, and excess culture medium is discharged into the first outlet reservoir through the first outlet port, providing the nutrients required for the growth of the intestinal epithelial cells.

[0091] Endothelial cells are added to the second inlet reservoir of the chip, entering and filling the second culture fluid channel through the second inlet port; this allows the endothelial cells to adhere to the hydrogel surface at the junction of the second culture fluid channel and the hydrogel channel; endothelial cell culture medium (culture fluid) is added to the second inlet reservoir of the chip, entering and filling the second culture fluid channel through the second inlet port, and excess culture medium is discharged into the second outlet reservoir through the second outlet port, providing the endothelial cells with the nutrients required for growth.

[0092] Liquid-driven methods can be pump-free gravity-driven or use external peristaltic pumps, syringe pumps, or other devices to drive liquid flow, enabling the liquid to provide the fluid shear force environment required for cell growth.

[0093] After successful cell culture, an intestinal-endothelial organ model was constructed for subsequent testing.

[0094] Implementation Method 2

[0095] The steps involved in three-dimensional cell culture using a microfluidic chip may include:

[0096] The second substrate is hydrophilically treated;

[0097] Before using the chip (in this scheme, a three-dimensional cell culture chip), the chip is sterilized.

[0098] When in use, add the hydrogel containing the mixed cells into the gel reservoir of the chip, and enter the hydrogel channel through the gel inlet. Due to the blocking effect of the liquid boundary, the hydrogel solution is confined in the hydrogel channel and does not flow into the other two culture fluid channels of the channel. Excess hydrogel flows out through the gel outlet and into the gel outlet reservoir.

[0099] After the hydrogel solidifies, it forms hydrogel strips.

[0100] Cell culture medium is added to the first and second inlet reservoirs of the chip, sequentially entering and filling the two culture fluid channels; excess hydrogel flows out through the outlet into the outlet reservoir; a membrane-free three-layer structure of culture medium-hydrogel-culture medium is formed in the chip.

[0101] Liquid-driven methods can be pump-free gravity-driven or use external peristaltic pumps, syringe pumps, or other devices to drive liquid flow, enabling the liquid to provide the fluid shear force environment required for cell growth.

[0102] Subsequent tests were conducted after successful cell culture.

[0103] In the accompanying drawings of this embodiment, the same or similar reference numerals correspond to the same or similar components. In the description of this application, it should be understood that if terms such as "upper," "lower," "left," and "right" 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, the terms used to describe positional relationships in the accompanying drawings are only for illustrative purposes and should not be construed as limiting this patent. For those skilled in the art, the specific meaning of the above terms can be understood according to the specific circumstances.

[0104] The above are merely preferred embodiments of this application and are not intended to limit this application. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of this application should be included within the protection scope of this application.

Claims

1. A microfluidic chip, characterized by, The microfluidic chip comprises: a main body having hydrogel channels and culture fluid channels which are spaced apart and partially connected; a liquid inlet storage structure having a hydrogel inlet storage pool and a liquid inlet storage pool, the hydrogel inlet storage pool being in communication with one end of the hydrogel channel, and the liquid inlet storage pool being in communication with one end of the culture fluid channel; a liquid outlet storage structure having a hydrogel outlet storage pool and a liquid outlet storage pool, the hydrogel outlet storage pool being in communication with the other end of the hydrogel channel, and the liquid outlet storage pool being in communication with the other end of the culture fluid channel; a liquid blocking structure arranged in the main body and located between the hydrogel channel and the culture fluid channel, a communication port being formed between the liquid blocking structure and the main body; wherein the hydrogel can enter the hydrogel channel through the hydrogel inlet storage pool and can be discharged from the hydrogel outlet storage pool, the hydrogel in the hydrogel channel being blocked by the liquid blocking structure and unable to flow into the culture fluid channel; the culture fluid can enter the culture fluid channel through the liquid inlet storage pool and can be discharged from the liquid outlet storage pool, and the culture fluid can exchange substances with the hydrogel through the communication port; the liquid inlet storage structure has a cylindrical liquid inlet storage space, which is divided into the hydrogel inlet storage pool and the liquid inlet storage pool with equal capacities; the liquid outlet storage structure has a cylindrical liquid outlet storage space, which is divided into the hydrogel outlet storage pool and the liquid outlet storage pool with equal capacities; the number of the hydrogel channels is one, and the number of the culture fluid channels is two, the hydrogel channel being located between the two culture fluid channels, and the liquid blocking structure being arranged between each culture fluid channel and the hydrogel channel; the liquid inlet storage structure includes three second liquid inlet partition plates, the three second liquid inlet partition plates being arranged in the liquid inlet storage space, one end of the three second liquid inlet partition plates being connected, and the other end being connected to the inner wall of the liquid inlet storage structure in a diverging manner, and the included angle between any two adjacent second liquid inlet partition plates being 120 degrees, so as to divide the cylindrical liquid inlet storage space into one hydrogel inlet storage pool and two liquid inlet storage pools with equal capacities; the liquid outlet storage structure includes three second liquid outlet partition plates, the three second liquid outlet partition plates being arranged in the liquid outlet storage space, one end of the three second liquid outlet partition plates being connected, and the other end being connected to the inner wall of the liquid outlet storage structure in a diverging manner, and the included angle between any two adjacent second liquid outlet partition plates being 120 degrees, so as to divide the cylindrical liquid outlet storage space into one hydrogel outlet storage pool and two liquid outlet storage pools with equal capacities; the length of the hydrogel channel is greater than the length of any culture fluid channel, and the lengths of the two culture fluid channels are equal, and the two ends of the hydrogel channel are protruding relative to the culture fluid channels; the hydrogel inlet storage pool is farther away from the liquid outlet storage structure than the two liquid inlet storage pools, so that the hydrogel inlet storage pool is in communication with one end of the hydrogel channel which is protruding relative to the culture fluid channels; The glue outlet reservoir is farther away from the liquid inlet reservoir structure than the two liquid outlet reservoirs, so that the glue outlet reservoir is in communication with the end of the hydrogel channel protruding relative to the culture fluid channel; The bottom of the glue inlet reservoir has a glue inlet port, and the flow cross section of the glue inlet port is smaller than that of the glue inlet reservoir; the bottom of the liquid inlet reservoir has a liquid inlet port, and the flow cross section of the liquid inlet port is smaller than that of the liquid inlet reservoir; The bottom of the glue outlet reservoir has a glue outlet port, and the flow cross section of the glue outlet port is smaller than that of the glue outlet reservoir; the bottom of the liquid outlet reservoir has a liquid outlet port, and the flow cross section of the liquid outlet port is smaller than that of the liquid outlet reservoir; The main body comprises: A first substrate having the hydrogel channel and the culture fluid channel; A second substrate having the glue inlet port, the liquid inlet port, the glue outlet port and the liquid outlet port; A third substrate having the liquid inlet reservoir structure and the liquid outlet reservoir structure; The liquid blocking structure comprises a first liquid blocking member and a second liquid blocking member arranged at intervals, and the hydrogel channel and the culture fluid channel are separated by the first liquid blocking member and the second liquid blocking member, the first liquid blocking member is arranged on the surface of the first substrate facing the second substrate, the second liquid blocking member is arranged on the surface of the second substrate facing the first substrate, and the communication port is formed between the first liquid blocking member and the second liquid blocking member.

2. The microfluidic chip of claim 1, wherein: The number of the hydrogel channel and the culture fluid channel is one; The liquid inlet reservoir structure comprises a first liquid inlet partition plate, and the first liquid inlet partition plate divides the liquid inlet reservoir space into the glue inlet reservoir and the liquid inlet reservoir with equal capacity; The liquid outlet reservoir structure further comprises a first liquid outlet partition plate, and the first liquid outlet partition plate divides the liquid outlet reservoir space into the glue outlet reservoir and the liquid outlet reservoir with equal capacity.

3. The microfluidic chip of claim 1, wherein, The side of the second substrate facing the first substrate is covered with a hydrophilic film or is subjected to hydrophilic surface treatment, or the entire second substrate is subjected to hydrophilic surface treatment.

4. The microfluidic chip of claim 1, wherein, The first liquid blocking member is at least one of a groove, a bar, a boss, a hydrophobic film and a grid; and the second liquid blocking member is at least one of a groove, a bar, a boss, a hydrophobic film and a grid.

5. The microfluidic chip of any one of claims 1-4, wherein: The width of the hydrogel channel is a, and 200 μm≤a≤600 μm; The width of the culture fluid channel is b, and 300 μm≤b≤700 μm; The height of the hydrogel channel and the culture fluid channel is h, and 200 μm≤h≤400 μm.

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