Microfluidic cell culture detection chip with functional factor steady-state function

Abstract

The invention relates to a micro-fluidic cell culture detection chip with a functional factor steady-state function. The micro-fluidic cell culture detection chip comprises a chip main body and a plurality of sealing membranes for being attached to the chip main body, the chip main body is provided with a centrifugal shaft and a plurality of culture detection cavity groups; the centrifugal shaft is used for being inserted into a matched centrifugal detection instrument and is positioned in the center of the chip main body; the culture detection cavity groups are circumferentially formed in the centrifugal shaft and are used for cell culture; the culture detection cavity group comprises a reaction section, a culture section, a detection section, a culture reaction channel and a transfer channel; a filling cavity for placing paraffin is formed in the side wall of the culture reaction channel, a floater for blocking the transfer channel is arranged in the transfer channel, and the floater can move under the action of gas generated in the cell culture process. Under the combined action of the filling paraffin and the floater, the culture section is sealed, and the interference effect of the external environment on cell culture is further reduced.

Description

Technical Field

[0001] The present application relates to the technical field of cell culture devices, and in particular to a microfluidic cell culture detection chip with a functional factor stabilization function. Background Art

[0002] Microfluidic chip technology integrates basic operating units such as sample preparation, reaction, separation, and detection in biological, chemical, and medical analytical processes onto micron-scale chips based on microstructure design. It uses the driving force generated by centrifugation to achieve complex experimental processes, such as cell evaluation. It has the characteristics of precise microenvironment regulation, low sample consumption, and high-throughput parallel analysis. Therefore, it has great application potential in cell evaluation experiments.

[0003] Cell evaluation tests are important tools in pharmacology and biomedical research, primarily used to assess the biological activity, mechanism of action, and safety of functional components. In existing technologies, traditional cell evaluation tests for functional components are subject to the challenges of complex and variable cell culture environments, coupled with the extremely unstable nature of functional factors such as flavonoids and polyphenols, which are susceptible to degradation due to environmental factors, seriously affecting the success rate of cell evaluation tests. Furthermore, the separation methods for functional components typically rely on centrifugation and manual operation, which present disadvantages such as complex operations, low processing efficiency, and difficulty in controlling experimental conditions. Furthermore, environmental pollution issues exist during cell chip culture, which can affect the cell evaluation of functional factors and other external environmental factors.

[0004] Therefore, it is necessary to design a microfluidic chip with the function of stabilizing functional factors, which can improve the stability of functional factor cell cultivation and reduce the impact of the external environment on cell cultivation. Summary of the Invention

[0005] In order to reduce the impact of the external environment on the cell culture process, the present application provides a microfluidic cell culture detection chip with the function of stabilizing functional factors, which adopts the following technical solutions: A microfluidic cell culture detection chip with a functional factor stabilization function comprises a chip body and several sealing membranes for being attached to the chip body; the chip body is provided with a centrifugal shaft located at the center of the chip body for being plugged into a matching centrifugal detection instrument, and several culture detection cavity groups circumferentially arranged on the centrifugal shaft and used for cell culture; the culture detection cavity group comprises a reaction section, a culture section, a detection section, a culture reaction channel and a transfer channel; the reaction section is used for mixed embedding of embedding reagents and reagents to be embedded, at which time the speed of the matching centrifugal detection instrument during the embedding process can be set to a second speed; the culture section is used for mixed culture of embedded reagents and cell fluid, at which time the culture process The rotation speed of the supporting centrifugal detection instrument can be set to a fourth rotation speed; the detection section is used to output the cultured product and perform detection, the culture reaction channel is used to connect the reaction section (13) and the culture section, and the rotation speed of the supporting centrifugal detection instrument for transferring the material from the reaction section to the culture section can be set to a third rotation speed; the transfer channel is used to connect the culture section and the detection section, and the rotation speed of the supporting centrifugal detection instrument for transferring the material from the culture section to the detection section can be set to a fifth rotation speed; a filling chamber for placing paraffin is provided on the side wall of the culture reaction channel, and a float is provided in the transfer channel to block the transfer channel, and the float can move under the driving action of the gas pressure generated during the cell culture process.

[0006] By adopting the above technical solution, during the cell culture process, under the action of the third speed of the supporting centrifugal detection instrument, the embedded reagents and cell culture fluid are transferred to the culture section for mixing. Then, by heating the chip body, the solid paraffin wax in the filling chamber is melted, becoming liquid and flowing in the culture reaction channel. After heating for a period of time, the heating is stopped, allowing the paraffin wax to solidify at room temperature, blocking the culture reaction channel, and making the reaction section and the culture section in a sealed state. A float is provided in the transfer channel so that the float seals the transfer channel. Under the combined action of the filling paraffin and the float, the culture section is sealed, further reducing the interference of the external environment on the cell culture. At the same time, based on the pressure changes mediated by CO2 and other gases generated during the cell culture process, the float shifts in the transfer channel, dynamically adjusting the internal pressure balance of the system, significantly reducing the risk of cell culture failure caused by pressure fluctuations.

[0007] Optionally, the reaction section includes a plurality of embedding reagent chambers for storing embedding reagents, a plurality of to-be-embedded chambers for storing to-be-embedded reagents, a reaction chamber for mixing embedding reagents and to-be-embedded reagents, an embedding reagent channel, and a to-be-embedded channel; the embedding reagent channel is used to connect the embedding reagent chamber and the reaction chamber, and the to-be-embedded channel is used to connect the to-be-embedded chamber and the reaction chamber. At this time, the speed of the supporting centrifugal detection instrument for transferring the reagents in the to-be-embedded chamber and the embedding reagent chamber to the reaction chamber can be set to a first speed; wherein The embedding reagent chamber is arranged on the side of the reaction chamber close to the centrifugal axis, and the chamber to be embedded is arranged on the side of the reaction chamber close to the centrifugal axis; the culture section includes a culture fluid chamber for placing cell culture fluid, a culture chamber, and a culture fluid channel; the culture chamber is used to mix the cell culture fluid and the embedding product in the reaction chamber and cultivate cells, and the culture fluid channel is used to connect the culture fluid chamber and the culture chamber; the culture fluid chamber is arranged on the side of the culture chamber close to the centrifugal axis; the culture reaction channel is curved.

[0008] By adopting the above technical solution, when it is necessary to mix the embedding reagent and the reagent to be embedded, at the first speed of the supporting centrifugal detection instrument, the embedding reagent enters the reaction chamber through the embedding reagent channel, and the reagent to be embedded enters the reaction chamber through the embedding channel. Since the culture reaction channel is bent, after the embedding reagent and the reagent to be embedded enter the reaction chamber, they will not enter the culture chamber through the culture reaction channel. After the embedding reagent and the reagent to be embedded are reacted, the speed of the supporting centrifugal detection instrument is adjusted to the second speed, so that the product and the cell culture fluid enter the culture chamber under the action of the second speed of the supporting centrifugal detection instrument. In the process of mixing the reagents, no manpower is required, and only the centrifugal speed of the supporting centrifugal detection instrument needs to be changed. The reagent mixing is more convenient. Since the culture reaction channel is set to a bent state, the next step of culture will not be carried out if the previous step has not been completed, further reducing the interference of the external environment on the cell culture. By delaying the flow rate of the reactants, the embedding droplets formed in the reaction section are stabilized, which can effectively improve the stability of the functional factor cell culture.

[0009] Optionally, a vent hole is provided on the side wall of the transfer channel for connecting the culture chamber to the outside world.

[0010] By adopting the above technical solution, when the gas generated during the cell culture process pushes the float to the position between the exhaust hole and the culture chamber, the culture process is in a closed state; when the float is pushed to the side of the exhaust hole away from the culture chamber, the culture process ends and the culture chamber is connected to the outside world. Under the action of siphoning and centrifugal force, the float and the product fluid can move together along the culture reaction channel to the detection section, reducing the influence of the float on fluid transfer.

[0011] Optionally, the transfer channel is further provided with a wavy bending section on a side of the exhaust hole away from the culture chamber.

[0012] By adopting the above technical solution, under the action of the fifth speed of the matching centrifugal detection instrument, when the cultured product is transferred to the detection section, the moving speed of the product is reduced through the bending section in the transfer channel, so that the product is more uniform after entering the detection section.

[0013] Optionally, the detection section includes several detection chambers for placing cell culture products and an output channel, and the output channel is used to connect the detection chamber and the transfer channel; the output channel is arranged on the side of the transfer channel away from the centrifugal axis, and the detection chamber is arranged on the side of the output channel away from the centrifugal axis; the output channel is arc-shaped, and the center of the circle coincides with the center of the centrifugal axis.

[0014] By adopting the above technical solution, during the output detection process of the final product, since the output channel and the centrifugal axis have the same center, the difference between the products in each detection chamber is reduced during the centrifugation process, further increasing the stability of the cell culture process.

[0015] Optionally, the detection section further includes a waste liquid tank, which is arranged on a side of the output channel away from the transfer channel and is communicated with the output channel.

[0016] By adopting the above technical solution, after the product enters the detection chamber, other liquids directly enter the waste liquid tank through the output channel, reducing contamination of the sample and further increasing the accuracy of the detection process.

[0017] Optionally, the detection section is further connected to a ventilation channel on a side of the waste liquid tank away from the output channel, and a ventilation hole is provided on a side of the ventilation channel away from the waste liquid tank.

[0018] By adopting the above technical solution, the vent holes can connect the culture and detection cavity group with the outside world, maintain stable air pressure, and reduce the probability of flow disturbance caused by gas compression.

[0019] Optionally, the depth of the reagent embedding channel is greater than the depth of the channel to be embedded.

[0020] By adopting the above technical solution, the reagent to be embedded is embedded by the embedding reagent when it passes into the reaction chamber from the embedding channel.

[0021] Optionally, a buffer zone is further provided in the reaction chamber, the buffer zone is connected to both the embedding reagent channel and the channel to be embedded, and the depth of the buffer zone is greater than that of the embedding reagent channel, and the depth of the buffer zone is less than that of the reaction chamber.

[0022] By adopting the above technical solution, when the reagents enter the reaction chamber, they are first discharged from the channel separately, then mixed in the buffer zone, and under the action of the buffer zone, form droplets, and then enter the reaction chamber, so that the mixing effect of the embedding reagent and the reagent to be embedded is better, and the reagent embedding is more uniform.

[0023] In summary, this application includes at least one of the following beneficial technical effects: During the cell cultivation process, under the action of the third speed of the matching centrifugal detection instrument, the embedded reagents and cell culture fluid are transferred to the culture section for mixing, and then the chip body is heated to melt the paraffin in the filling chamber, which becomes liquid and flows in the culture reaction channel; after heating for a period of time, the heating is stopped to allow the paraffin to solidify at room temperature, and the culture reaction channel is sealed, so that the reaction section and the culture section are in a sealed state; a float is provided in the transfer channel so that the float seals the transfer channel. Under the joint action of the filling paraffin and the float, the culture section is sealed, further reducing the interference of the external environment on the cell culture. At the same time, because the float can move in the transfer channel through the gas generated during the cell culture process, the probability of cultivation failure due to air pressure factors during the experiment is reduced; When the gas generated during the cell culture process pushes the float between the vent and the culture chamber, the culture process is closed. When the float is pushed to the side of the vent away from the culture chamber, the culture process ends and the culture chamber is connected to the outside world. Under the action of siphoning and centrifugal force, the float and the product fluid can move together along the culture reaction channel to the detection section, reducing the influence of the float on fluid transfer. When the reagents enter the reaction chamber, they are first discharged from the channel, then mixed in the buffer zone, and under the action of the buffer zone, they form droplets, and then enter the reaction chamber, so that the mixing effect of the embedding reagent and the reagent to be embedded is better, and the reagent embedding is more uniform. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 This is a schematic diagram of the overall structure of a microfluidic cell culture detection chip with the function of stabilizing functional factors.

[0025] Figure 2 It is a schematic diagram of the structure of the culture detection cavity group.

[0026] Figure 3 yes Figure 2 Enlarged view of point A in the middle.

[0027] Explanation of the accompanying drawings: 1. Chip body; 11. Centrifugal shaft; 12. Culture detection cavity group; 13. Reaction section; 131. Embedding reagent chamber; 132. Chamber to be embedded; 133. Reaction chamber; 1331. Buffer zone; 134. Embedding reagent channel; 135. Channel to be embedded; 14. Culture section; 141. Culture fluid chamber; 142. Culture chamber; 143. Culture fluid channel; 15. Detection section; 151. Output channel; 152. Detection chamber; 153. Waste liquid tank; 154. Vent channel; 155. Vent hole; 16. Culture reaction channel; 161. Filling chamber; 17. Transfer channel; 171. Moving section; 1711. Float; 172. Bending section; 173. Transition section; 1731. Exhaust hole; 2. Sealing diaphragm; 21. First membrane hole; 22. Second membrane hole. DETAILED DESCRIPTION

[0028] The following is combined with Figure 1-2 This application is described in further detail.

[0029] The embodiments of the present application disclose a microfluidic cell culture detection chip with the function of stabilizing functional factors.

[0030] Reference Figure 1 A microfluidic cell culture chip includes a chip body 1 and a plurality of sealing membranes 2 for fitting the chip body 1. The chip body 1 is provided with a centrifugal shaft 11 for being plugged into a matching centrifugal detection instrument and a plurality of culture detection cavity groups 12 for cell culture, which are provided on one side close to the sealing membrane 2. In this embodiment, the chip body 1 is in the shape of a flat disc as a whole and is provided with two groups of culture detection cavity groups 12, and the two groups of culture detection cavity groups 12 are arranged on both sides of the chip body 1. The number of sealing membranes 2 corresponds to the number of culture detection cavity groups 12, and the culture detection cavity groups 12 are covered to reduce the probability of reactants in the chip body 1 being thrown out during the centrifugation process.

[0031] Reference Figure 1 and Figure 2The culture detection cavity group 12 includes a reaction section 13, a culture section 14, a detection section 15, a culture reaction channel 16, and a transfer channel 17. In this embodiment, the reaction section 13 is used for embedding the reagent to be embedded and the embedding reagent for embedding reaction, the culture section 14 is used for mixed culture of the reactants in the reaction section 13 and the cell fluid, the detection section 15 is used to detect the culture products of the culture section 14, the culture reaction channel 16 is used to connect the reaction section 13 and the culture section 14, and the transfer channel 17 is used to connect the culture section 14 and the detection section 15.

[0032] In order to facilitate the transfer of materials and to suit the reaction of materials, the supporting centrifugal detection instrument is also provided with different speeds. The speed for transferring the embedding reagent and the reagent to be embedded is the first speed, the speed for the embedding reaction process in the reaction section 13 is the second speed, the speed for transferring the product of the reaction section 13 to the culture section 14 is the third speed, the speed for the culture process in the culture section 14 is the fourth speed, and the speed for transferring the product of the culture section 14 to the detection section 15 is the fifth speed.

[0033] Reference Figure 2 and Figure 3 The reaction section 13 includes a plurality of embedding reagent chambers 131 for storing embedding reagents, a plurality of embedding chambers 132 for storing reagents to be embedded, a reaction chamber 133 for reacting the reagents to be embedded and the embedding reagents, an embedding reagent channel 134, and a channel 135 for embedding. The embedding reagent channel 134 is used to connect the embedding reagent chamber 131 and the reaction chamber 133, and the channel 135 is used to connect the embedding chamber 132 and the reaction chamber 133. In this embodiment, the reaction section 13 has two embedding reagent chambers 131 and one embedding chamber 132, and the embedding chamber 132 is arranged between the two embedding reagent chambers 131. The embedding reagent chamber 131 and the embedding chamber 132 are both located on the side of the reaction chamber 133 close to the centrifugal shaft 11. When the reagent to be embedded and the embedding reagent need to be transported to the reaction chamber 133, the operator needs to place the reagent to be embedded in the embedding chamber 132 and the embedding reagent in the embedding reagent chamber 131, and then attach the sealing membrane 2 to each. Then, the first rotational speed is started, and at the first rotational speed, the reagents enter the reaction chamber 133 to carry out the embedding reaction. To enhance the reaction effect of the embedding reagent and the reagent to be embedded, the embedding reagent channel 134 is deeper than the embedding channel 135, so that the reagent to be embedded is embedded by the embedding reagent when it passes from the embedding channel 135 into the reaction chamber 133.

[0034] Reference Figure 2 and 3A buffer zone 1331 is further provided in the reaction chamber 133. In this embodiment, the buffer zone 1331 is provided on a side of the reaction chamber 133 close to the centrifugal shaft 11 and is connected to both the embedding reagent channel 134 and the channel to be embedded 135. In order to increase the buffering effect on the reagent, the depth of the buffer zone 1331 is greater than the depth of the embedding reagent channel 134 but less than the depth of the reaction chamber 133. During the centrifugation process, when the reagent enters the reaction chamber 133, it first forms droplets under the action of the buffer zone 1331 and then enters the reaction chamber 133; when all the reagents have entered the reaction chamber 133, the speed is switched to the second speed to carry out the embedding reaction.

[0035] Reference Figure 2 and 3 One end of the culture reaction channel 16 is connected to the side of the reaction chamber 133 away from the centrifugal shaft 11, and the other end is connected to the culture section 14. A plurality of filling chambers 161 are provided on the side wall of the culture reaction channel 16. In this embodiment, the culture reaction channel 16 is curved, and the filling chambers 161 are used to place solid paraffin wax, which is used to seal the culture reaction channel 16. After the reagent reaction in the reaction chamber 133 is completed, the speed is changed to the third speed, and the product is passed into the culture section 14 through the culture reaction channel 16. In order to prevent the external environment from affecting the reagents in the culture section 14 through the culture reaction channel 16, after the product passes through the culture reaction channel 16, the paraffin is heated to 50-70°C. When the paraffin is in a liquid state and flows into the culture reaction channel 16, the heating is stopped to solidify the paraffin, thereby sealing the culture reaction channel 16 and reducing the probability of the external environment contaminating the reagents in the culture section 14 through the culture reaction channel 16. The heating method can be to directly heat the chip body 1 through a centrifugal device, or to provide a heating wire in the filling chamber 161. In the case of the heating wire, a metal wire that can undergo a large expansion deformation when heated to 50-60°C, such as a nickel-titanium alloy wire, can be provided in the filling chamber 161, and then a control component for controlling the heating of the metal wire is provided on the chip body 1, so that the metal wire melts the paraffin by heating and pushes the paraffin out of the filling chamber 161 by expansion. Compared with the method of directly heating the chip body, this heating method is more convenient to control the temperature and is more stable, reducing the cultivation uncertainty caused by heating the cell culture medium.

[0036] Reference Figure 2 and 3The culture section 14 includes a culture fluid chamber 141, a culture chamber 142 and a culture fluid channel 143. The culture fluid chamber 141 is used to place a cell culture fluid containing cells, the culture chamber 142 is used for culturing cells, and the culture fluid channel 143 is used to connect the culture fluid chamber 141 and the culture chamber 142. In this embodiment, the culture fluid chamber 141 is located on the side of the culture chamber 142 close to the centrifugal shaft 11. When cells need to be cultured, the operator places the cell culture fluid in the culture fluid chamber 141 at the beginning of the cultivation, and at the third speed, it enters the culture chamber 142 together with the reaction chamber 133, and is combined with the product of the reaction chamber 133 for mixing, and the product in the reaction chamber 133 and the cell culture fluid all enter the culture chamber 142, and the rotation speed is changed to the fourth speed, and the culture is carried out at the fourth speed.

[0037] Reference Figure 2 and 3 One end of the transfer channel 17 is connected to the side of the culture chamber 142 away from the centrifugal shaft 11. The transfer channel 17 includes a moving section 171, a bending section 172 and a transition section 173. In the present embodiment, the transition section 173 is located between the moving section 171 and the bending section 172, and the transition section 173 has a bend. The transition section 173 is provided with an exhaust hole 1731 on the side wall of the inflection point close to the centrifugal shaft 11 for communicating with the outside world. The moving section 171 is located on the side of the transition section 173 close to the culture chamber 142, and a float 1711 is provided in the moving section 171 that can be pushed by the gas generated during the cell cultivation process. Since the cell culture process needs to be isolated from the influence of the external environment, and gas is generated during the culture process, during the culture stage, the float 1711 is located in the moving section 171 and moves toward the exhaust hole 1731 during the culture process. After a certain period of incubation, float 1711 moves to vent 1731, connecting culture chamber 142 to the outside world through vent 1731, completing cell culture. The rotation speed then shifts to the fifth rotational speed, whereupon the product and float 1711 are transported along transfer channel 17 to detection section 15. As the liquid passes through bend 172, its multiple wavy turns slow its flow rate, resulting in a more even distribution of substances within the liquid upon entering detection section 15.

[0038] Reference Figure 2 and Figure 3The detection section 15 includes an output channel 151 connected to the transfer channel 17, several detection chambers 152 defined in the sidewalls of the output channel 151, a waste liquid trough 153 located at the end of the output channel 151 remote from the detection section 15, a ventilation channel 154 located at the end of the waste liquid trough 153 remote from the output channel 151, and a ventilation hole 155 located on the side of the air outlet channel remote from the waste liquid trough 153. In this embodiment, five detection chambers 152 are provided. To ensure uniform liquid composition within each detection chamber 152, the output channel 151 is arc-shaped, with its center coinciding with the center of the centrifugal shaft 11. The detection chambers 152 are located on the side of the output channel 151 remote from the centrifugal shaft 11. Under the influence of the fifth rotational speed of the accompanying centrifugal detection instrument, the liquid to be tested enters the output channel 151 through the output transfer channel 17 and is evenly distributed into the multiple detection chambers 152. Excess liquid and the float 1711 carried by it enter the waste liquid trough 153 for recovery. Because different test substances are required in each test chamber 152 during the testing process, the five test chambers 152 are circumferentially divided into a first test chamber 152A, a second test chamber 152B, a third test chamber 152C, a fourth test chamber 152D, and a fifth test chamber 152E. The vents 155 and vent channels 154 connect the interior of the culture test chamber group 12 with the outside world, maintaining stable air pressure and reducing the possibility of flow disturbances caused by gas compression.

[0039] Reference Figure 1 and Figure 3 The sealing membrane 2 is fan-shaped. In this embodiment, the sealing membrane 2 is used to fit over the chip body 1 and provide a seal. The sealing membrane 2 includes a first membrane hole 21 that corresponds one-to-one with the exhaust holes 1731 and has a larger diameter than the exhaust holes 1731, and a second membrane hole 22 that corresponds one-to-one with the vent holes 155 and has a larger diameter than the vent holes 155. This allows the culture and detection cavity group 12 to communicate with the outside world during use, balancing the internal and external air pressures.

[0040] The implementation principle of a microfluidic cell culture detection chip with a functional factor stabilization function in the embodiment of the present application is as follows: 1. Install the chip body 1 on the supporting centrifugal detection instrument, then inject the embedding reagent into the embedding reagent chamber 131, the reagent to be embedded into the embedding chamber 132, and the cell culture medium into the culture medium chamber 141. Then attach the sealing membrane 2. During the attachment process, it is important to note that the first membrane hole 21 on the sealing membrane 2 corresponds to the exhaust hole 1731, and the second membrane hole 22 corresponds to the vent hole 155; 2. Start the supporting centrifugal detection instrument and switch it to the first speed, so that the reagent to be embedded and the embedding reagent enter the reaction chamber 133 and are mixed; 3. Adjust the speed of the supporting centrifugal detection instrument to the second speed so that the embedded reagent and the embedding reagent are embedded. After the embedding is completed, change the speed of the supporting centrifugal detection instrument to the third speed so that the embedded product passes through the culture reaction channel 16 into the culture chamber 142, and the cell culture fluid also enters the culture chamber 142 through the culture fluid channel 143; 4. After the two are mixed, the chip body 1 is heated appropriately, and the temperature is controlled between 50-70°C, so that the solid paraffin in the filling chamber 161 is liquefied and flows into the culture reaction channel 16; 5. After heating for a period of time, the temperature is stopped and allowed to drop, the paraffin wax solidifies, the culture reaction channel 16 is sealed, and the speed is switched to the fourth speed to start cell culture; 6. During the cell culture process, the transfer channel 17 is also blocked due to the presence of the float 1711 in the transfer channel 17. During the culture process, gas is generated, pushing the float 1711 to move in the transfer channel 17. 7. After a period of culture, the float 1711 is pushed to the exhaust hole 1731, and the culture chamber 142 is connected to the outside world. At this time, the cell culture is completed; 8. Adjust the speed of the supporting centrifugal detection instrument to the fifth speed, so that the product passes through the transfer channel 17 and the output channel 151 under the action of the supporting centrifugal detection instrument and enters the detection chamber 152. The remaining waste liquid and the float 1711 enter the waste liquid tank 153, and then the instrument is used to detect the products in different detection chambers 152.

[0041] The above are all preferred embodiments of the present application, and are not intended to limit the scope of protection of the present application. Therefore, any equivalent changes made based on the structure, shape, and principle of the present application should be included in the scope of protection of the present application.

Claims

1. A microfluidic cell culture detection chip with a functional factor stabilization function, characterized in that: The invention comprises a chip body (1) and a plurality of sealing membranes (2) for being attached to the chip body (1); the chip body (1) is provided with a centrifugal shaft (11) for being plugged into a matching centrifugal detection instrument and located at the center of the chip body (1), and a plurality of culture detection cavity groups (12) circumferentially provided on the centrifugal shaft (11) and used for cell culture; the culture detection cavity group (12) comprises a reaction section (13), a culture section (14), a detection section (15), a culture reaction channel (16) and a transfer channel (17); the reaction section (13) is used for mixed embedding of embedding reagents and reagents to be embedded, at which time the rotation speed of the matching centrifugal detection instrument during the embedding process can be set to the second rotation speed; the culture section (14) is used for mixed culture of embedded reagents and cell fluid, at which time the rotation speed of the matching centrifugal detection instrument during the cultivation process can be set to the fourth rotation speed. speed; the detection section (15) is used to output the cultured product and perform detection, the culture reaction channel (16) is used to connect the reaction section (13) and the culture section (14), at which time the rotation speed of the supporting centrifugal detection instrument for transferring the material from the reaction section (13) to the culture section (14) can be set to a third rotation speed; the transfer channel (17) is used to connect the culture section (14) and the detection section (15), at which time the rotation speed of the supporting centrifugal detection instrument for transferring the material from the culture section (14) to the detection section (15) can be set to a fifth rotation speed; a filling chamber (161) for placing paraffin is opened on the side wall of the culture reaction channel (16), and a float (1711) is provided in the transfer channel (17) to block the transfer channel (17), and the float (1711) can move under the driving action of the gas pressure generated during the cell culture process.

2. The microfluidic cell culture detection chip with the function of stabilizing functional factors according to claim 1, characterized in that: The reaction section (13) includes a plurality of embedding reagent chambers (131) for storing embedding reagents, a plurality of embedding chambers (132) for storing reagents to be embedded, a reaction chamber (133) for mixing embedding reagents and reagents to be embedded, an embedding reagent channel (134) and a channel to be embedded (135); the embedding reagent channel (134) is used to connect the embedding reagent chamber (131) and the reaction chamber (133), and the channel to be embedded (135) is used to connect the chamber to be embedded (132) and the reaction chamber (133). At this time, the speed of the supporting centrifugal detection instrument for transferring the reagents in the chamber to be embedded (132) and the embedding reagent chamber (131) to the reaction chamber (133) can be set to a first speed; wherein the embedding reagent chamber (133) 31) is arranged on the side of the reaction chamber (133) close to the centrifugal shaft (11), and the chamber to be embedded (132) is arranged on the side of the reaction chamber (133) close to the centrifugal shaft (11); the culture section (14) includes a culture fluid chamber (141) for placing cell culture fluid, a culture chamber (142), and a culture fluid channel (143); the culture chamber (142) is used to mix the cell culture fluid and the embedded product in the reaction chamber (133) and cultivate cells, and the culture fluid channel (143) is used to connect the culture fluid chamber (141) and the culture chamber (142); the culture fluid chamber (141) is arranged on the side of the culture chamber (142) close to the centrifugal shaft (11); the culture reaction channel (16) is bent.

3. The microfluidic cell culture detection chip with the function of stabilizing functional factors according to claim 2, characterized in that: An exhaust hole (1731) is provided on the side wall of the transfer channel (17) for allowing the culture chamber (142) to communicate with the outside world.

4. The microfluidic cell culture detection chip with the function of stabilizing functional factors according to claim 3, characterized in that: The transfer channel (17) is further provided with a wavy bending section (172) on the side of the exhaust hole (1731) away from the culture chamber (142).

5. The microfluidic cell culture detection chip with the function of stabilizing functional factors according to claim 1, characterized in that: The detection section (15) comprises a plurality of detection chambers (152) for placing cell culture products and an output channel (151), wherein the output channel (151) is used to connect the detection chambers (152) and the transfer channel (17); the output channel (151) is arranged on a side of the transfer channel (17) away from the centrifugal shaft (11), and the detection chamber (152) is arranged on a side of the output channel (151) away from the centrifugal shaft (11); the output channel (151) is in an arc shape, and the center of the circle coincides with the center of the centrifugal shaft (11).

6. The microfluidic cell culture detection chip with the function of stabilizing functional factors according to claim 5, characterized in that: The detection section (15) further comprises a waste liquid tank (153), which is arranged on a side of the output channel (151) away from the transfer channel (17) and is communicated with the output channel (151).

7. The microfluidic cell culture detection chip with the function of stabilizing functional factors according to claim 6, characterized in that: The detection section (15) is further connected to a ventilation channel (154) on the side of the waste liquid tank (153) away from the output channel (151), and a ventilation hole (155) is provided on the side of the ventilation channel (154) away from the waste liquid tank (153).

8. The microfluidic cell culture detection chip with the function of stabilizing functional factors according to claim 2, characterized in that: The depth of the reagent embedding channel (134) is greater than the depth of the channel to be embedded (135).

9. The microfluidic cell culture detection chip with the function of stabilizing functional factors according to claim 2, characterized in that: A buffer zone (1331) is further provided in the reaction chamber (133), and the buffer zone (1331) is connected to both the embedding reagent channel (134) and the channel to be embedded (135), and the depth of the buffer zone (1331) is greater than that of the embedding reagent channel (134), and the depth of the buffer zone (1331) is less than that of the reaction chamber (133).

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