A disc type micro-fluidic chip and sealing method

Abstract

The application discloses a disc type micro-fluidic chip and a sealing method, relates to the field of micro-fluidics, and solves the problems of poor sealing performance and complicated operation of the existing disc type micro-fluidic chip. The disc type micro-fluidic chip comprises 16 independent reaction detection structures which are uniformly distributed on the circumference of the disc type micro-fluidic chip. Each reaction detection structure comprises a V-shaped sample chamber, a horseshoe-shaped reagent chamber and a circular detection chamber. The V-shaped sample chambers are connected with each other and located on the side close to the center of the disc type micro-fluidic chip. Sample injection holes and air holes are arranged at the two ends of the V-shaped sample chambers. The horseshoe-shaped reagent chambers are independent of each other and located in the middle of the disc type micro-fluidic chip. Each horseshoe-shaped reagent chamber is provided with two liquid injection holes and air holes. The circular detection chambers are independent of each other and located on the outer periphery of the disc type micro-fluidic chip and used for mixing and reacting the sample and the reagent. The application provides two sealing modes of the disc type micro-fluidic chip and is also suitable for application fields such as nucleic acid detection.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of microfluidic technology, and in particular to a disc-type microfluidic chip and a sealing method. BACKGROUND

[0002] Microfluidic PCR chips can integrate sample mixing, thermal cycling amplification, and product detection steps required for PCR reactions on a small chip platform by integrating microchannels, microreaction chambers, and other structures on the chip. This integrated design not only significantly reduces the amount of reagents used, reduces detection costs, but also improves reaction efficiency and sensitivity, enabling nucleic acid detection to be completed in a shorter time. Microfluidic PCR chips also have the characteristics of miniaturization and portability, which can be easily applied to on-site detection, bedside diagnosis, and other scenarios, meeting the urgent needs of modern medicine for rapid, accurate, and convenient detection methods.

[0003] In the development history of microfluidic PCR chips, disc-type microfluidic chips have gradually emerged as a research hotspot. Disc-type microfluidic chips use centrifugal force as the main method of fluid driving, and achieve the flow and distribution of samples and reagents in microchannels by rotating the chip. This driving method does not require additional external pumps or valves and other complex fluid driving equipment, simplifying the structure and operation process of the chip, reducing equipment costs and operation difficulty, and providing strong support for the commercialization and popularization of microfluidic PCR chips. Disc-type microfluidic chips can also achieve the integration of multiple channels and multiple reaction units, allowing parallel detection of multiple samples or multi-index analysis of a single sample, greatly improving the detection throughput and efficiency.

[0004] Although disc-type microfluidic chips have shown many advantages in the field of nucleic acid detection, they still face some challenges in practical application. For example, when performing multiple sample amplification, how to effectively avoid cross contamination is a key problem that needs to be solved. Due to the complex structure of the microchannels of the disc-type chip, leakage or residue may occur during the flow and reaction of the sample in the microchannels, leading to cross contamination between different samples and affecting the accuracy and reliability of the detection results. The existing disc-type microfluidic chip sampling method is often complex, requiring precise control of the sampling amount and speed, which is technically demanding for the operator and increases the risk of operational errors. In addition, the sealing performance of the chip is also an important factor affecting the detection results. If the chip is not sealed tightly, water evaporation and gas leakage may occur in the reaction system, affecting the PCR reaction and the accuracy of the detection results.

[0005] A centrifugal microfluidic chip is disclosed in Chinese Patent Publication No. CN111135879A, which relates to a centrifugal microfluidic chip and a sample processing method. The centrifugal microfluidic chip includes a liquid inlet hole, an equal division hole, a first siphon pipe, a sample pretreatment area, a second siphon pipe, a reaction hole, and a waste liquid pool connected in sequence. The sample pretreatment area is also connected to an exhaust hole. The equal division hole includes two cavities with equal volumes to achieve equal division of the sample; each reaction hole has an equal volume to achieve secondary division of the sample; and the sample pretreatment area and each reaction hole are loaded with different reaction reagents as needed to achieve multiple detection applications, or loaded with the same reaction reagent to achieve parallel detection of the same sample. However, the centrifugal microfluidic chip also has some limitations that cannot be ignored. First, the number of channels on the chip is too small, which is a significant problem. In the face of a large number of sample application scenarios, too few channels mean that the number of samples that can be processed at a time is limited. This results in the need for repeated operations when processing a large number of samples, consuming a lot of time and labor costs, and seriously affecting the overall detection efficiency. Second, the sealing problem of the chip also needs to be solved. During the sample reaction process, due to the insufficient sealing of the chip, aerosol pollution problems are easily caused. Aerosol refers to small particles suspended in the gas, and various aerosols may be produced during the sample reaction process. If the chip is not tightly sealed, these aerosols will leak outside the chip, not only causing pollution to the experimental environment, but also possibly affecting the detection results of other samples, and even posing a potential threat to the health of the operator. Moreover, aerosols can also spread between different reaction chambers inside the chip, interfering with the normal reaction process, causing the detection results to deviate, and reducing the reliability and accuracy of the experiment.

[0006] To solve the above problems, a PCR disc microfluidic chip with high-efficiency anti-pollution, multi-channel, rapid reaction, convenient sample loading, and high sealing performance, as well as a sealing method thereof, is developed. This not only promotes the development of microfluidic technology, but also plays an important role in clinical diagnosis, public health safety, biopharmaceuticals, and other fields, has a broad market prospect and far-reaching social value. SUMMARY

[0007] The present application is to solve the problems of poor sealing performance and complex operation of the disc microfluidic chip in the prior art.

[0008] To solve the above technical problems, the present application is realized by the following technical scheme:

[0009] Solution 1: The present invention proposes a disc-type microfluidic chip, which includes 16 reaction detection modules, which are evenly distributed on the circumference of the disc-type microfluidic chip. Each reaction detection module includes a chamber and a channel, specifically including a first ventilation hole, a circular detection chamber, a second ventilation hole, a horseshoe-shaped reagent chamber, a sealing valve, an injection hole, a first capillary valve, a second capillary valve, a V-shaped sample chamber, and a sample addition hole;

[0010] Each V-shaped sample chamber is connected to each other. The V-shaped sample chamber is arranged on one side of the disc-shaped microfluidic chip close to the center of the circle. The terminal positions on both sides of the V-shaped sample chamber are respectively provided with a sample addition hole or a vent hole. If one terminal is a sample addition hole, the other terminal is regarded as a vent hole.

[0011] The horseshoe-shaped reagent chamber is located in the middle of the disc-type microfluidic chip, and each horseshoe-shaped reagent chamber is provided with two injection holes or vents, where one end is the injection hole and the other end is the vent;

[0012] The circular detection chamber is located on the periphery of the disc-type microfluidic chip and is used to mix the sample and the reagent to generate a reaction.

[0013] Furthermore, a preferred embodiment is provided, wherein the disc-type microfluidic chip further comprises positioning holes and positioning pins, and the inner ring of the disc-type microfluidic chip is provided with positioning pins, and the outer ring is provided with positioning holes.

[0014] Furthermore, a preferred embodiment is provided, wherein the V-shaped sample chamber is connected to the horseshoe-shaped reagent chamber via a first capillary valve.

[0015] Furthermore, a preferred embodiment is provided, wherein the horseshoe-shaped reagent chamber is connected to the circular detection chamber via a second capillary valve.

[0016] Furthermore, a preferred embodiment is provided, wherein the disc-type microfluidic chip also includes an annular sealing film and a circular sealing film, wherein the annular sealing film is used to seal the sample addition hole or the vent hole on the horseshoe-shaped reagent chamber, and the circular sealing film is used to seal the two injection holes or the vent holes on the V-shaped sample chamber.

[0017] Furthermore, a preferred embodiment is provided, wherein the disk-type microfluidic chip further comprises a sealing valve, and the sealing valve is used to seal the second capillary valve.

[0018] Furthermore, a preferred embodiment is provided, wherein the disc-type microfluidic chip is provided with 16 first ventilation holes and 16 second ventilation holes, and only the disc-type microfluidic chip containing the sealing valve has the second ventilation holes.

[0019] Scheme two, a sealing method of a disc-shaped microfluidic chip, the method is realized based on the disc-shaped microfluidic chip in any one of the schemes one, the method comprises the following steps:

[0020] S1. In the state that the annular sealing rubber sheet tightly seals the liquid injection hole or the air hole on the horseshoe-shaped reagent chamber, the gun head of the pipette is aligned with the sample injection hole on the V-shaped sample chamber, and the sample diluent is injected into the V-shaped sample chamber;

[0021] S2. The sample injection hole on the V-shaped sample chamber is sealed with a circular sealing rubber sheet, and after the sealing is completed, the disc-shaped microfluidic chip is placed in a centrifuge, the centrifuge is started to apply a centrifugal force, and the sample diluent is successfully introduced into the circular detection chamber;

[0022] S3. After the sample transfer is completed, the annular sealing rubber sheet is removed, the annular sealing rubber sheet is removed, the pipette is picked up again, the reagents required for the reaction are sequentially sucked, the gun head of the pipette is inserted into the horseshoe-shaped reagent chamber through the liquid injection hole or the air hole, and the reagents are added to the horseshoe-shaped reagent chamber;

[0023] S4. After the reagent addition is completed, the disc-shaped microfluidic chip is placed in a centrifuge, the second capillary valve is moved to the circular detection chamber, and the reaction reagent enters the circular detection chamber and mixes with the sample diluent introduced in S3;

[0024] S5. After the sample and the reagent are introduced into the circular detection chamber, the reaction solution is sealed.

[0025] Further, a preferred embodiment is provided, the sealing of the reaction solution in S5 comprises a sealing valve sealing and a physical barrier layer method,

[0026] The sealing valve sealing method is:

[0027] S1.1. A clean pipette ready for use is provided with an appropriate gun head, an appropriate amount of ultraviolet curing glue is sucked, the gun head of the pipette is inserted into the sealing valve, the ultraviolet curing glue is injected into the sealing valve by pushing the push rod of the pipette, after the injection is completed, the chip is stably placed in a centrifuge, and the centrifugation time is set to 1 minute for quickly and uniformly filling the second capillary valve;

[0028] S1.2. After the centrifugation is completed, the disc-shaped microfluidic chip is taken out, the pen end of the ultraviolet irradiation pen is aligned with the second capillary valve, the ultraviolet curing glue in the second capillary valve is uniformly irradiated from the starting end of the second capillary valve;

[0029] S1.3, the original liquid ultraviolet curing adhesive gradually occurs crosslinking reaction, gradually changes from a flow state to a solid state, forms a solid and well-sealed adhesive structure, realizes sealing of the second capillary valve, and after sealing is completed, the disc microfluidic chip is taken out of the centrifuge and placed on a desktop.

[0030] Further, a preferred embodiment is provided, which comprises the following steps:

[0031] S2.1, the removed annular sealing film is re-covered and attached to the liquid injection hole of the horseshoe-shaped reagent chamber, and a physical barrier layer is formed by using the adhesion or pressure attachment of the sealing film itself;

[0032] S2.2, after sealing the liquid injection hole of the horseshoe-shaped reagent chamber, the ultraviolet curing adhesive is taken out using a pipette and slowly injected through the sample injection hole of the V-shaped sample chamber, and during the injection process, the chamber state needs to be continuously observed until the ultraviolet curing adhesive completely fills the V-shaped sample chamber, forming a convex higher than the plane of the sample injection hole;

[0033] S2.3, after the injection is completed, the ultraviolet lamp located at the center of the chip is turned on for 60 seconds of irradiation and curing, and the adhesive gradually changes from a liquid state to a solid state, finally forming a sealing layer on the surface of the sample injection hole.

[0034] The present application has the advantages of:

[0035] The disc microfluidic chip structure and the two sealing methods of the disc microfluidic chip and the sealing method according to the present application have obvious advantages. On the structure of the disc microfluidic chip, multiple independent reaction detection structures can be used to process multiple samples in parallel, improving the detection efficiency. The two sealing methods according to the present application, the sealing valve seals the channel by ultraviolet curing adhesive, blocking aerosol pollution, but each channel needs to be sealed one by one during irradiation and curing; without a sealing valve, the ultraviolet curing adhesive cooperates with the traditional mechanical sealing, but the liquid injection hole needs to be sealed by the traditional sealing method. The two sealing methods are flexible and suitable for different experimental scenarios. Both of them can effectively ensure the sealing of the reaction environment and improve the accuracy and reliability of nucleic acid amplification detection.

[0036] The present application is also applicable to the field of nucleic acid detection. BRIEF DESCRIPTION OF DRAWINGS

[0037] Figure 1 A top view of a disc microfluidic chip containing a sealing valve according to the first embodiment.

[0038] Figure 2 A bottom exploded isometric view of a disc microfluidic chip containing a sealing valve according to the first embodiment.

[0039] Figure 3A plan view of a disc-shaped microfluidic chip without a sealing valve according to embodiment one.

[0040] Figure 4 A bottom exploded isometric view of a disc-shaped microfluidic chip without a sealing valve according to embodiment one.

[0041] Figure 5 An enlarged schematic view of a flow channel of a disc-shaped microfluidic chip according to embodiment one.

[0042] Figure 6 An operation process schematic view of a disc-shaped microfluidic chip with a sealing valve according to embodiment one.

[0043] Figure 7 An operation process schematic view of a disc-shaped microfluidic chip without a sealing valve according to embodiment one.

[0044] Figure 8 A flow chart of a sealing method of a disc-shaped microfluidic chip according to embodiment eight.

[0045] Wherein, the first vent hole 1, the circular detection chamber 2, the second vent hole 3, the horseshoe-shaped reagent chamber 4, the sealing valve 5, the liquid injection hole 6, the first capillary valve 7, the second capillary valve 8, the V-shaped sample chamber 9, the sample injection hole 10, and the bottom substrate 11. DETAILED DESCRIPTION

[0046] In order to make the objects, technical solutions and advantages of the embodiments of the present application clearer, the technical solutions in the embodiments of the present application will be described clearly and completely below with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are some but not all of the embodiments of the present application.

[0047] Embodiment one, see Figures 1 to 8As shown, the present embodiment proposes a disc-shaped microfluidic chip, which includes 16 independent reaction detection structures. The 16 reaction detection structures are evenly distributed on the circumference of the disc-shaped microfluidic chip, which can make full use of the circumferential space of the disc-shaped chip and ensure that each reaction detection structure can operate independently and efficiently. Each reaction detection structure is composed of specific chambers and channels, specifically including V-shaped sample chambers 9, horseshoe-shaped reagent chambers 4, circular detection chambers 2, and first capillary valves 7 and second capillary valves 8. Each V-shaped sample chamber 9 is connected to each other and arranged on the side of the disc-shaped microfluidic chip close to the center. At the terminal position on both sides of the V-shaped sample chamber 9, a sample hole 10 and a vent hole are arranged. The two holes have a common function, in actual operation, one end can be used to add sample liquid, and the other end can be used to exhaust air to ensure smooth sample adding process. Each horseshoe-shaped reagent chamber 4 is independent of each other, which is located in the middle of the disc-shaped microfluidic chip. Each horseshoe-shaped reagent chamber 4 is provided with two liquid injection holes 6 and a vent hole, and the two holes also have a common function, one end can be used to inject reagent liquid, and the other end can be used to exhaust air to ensure smooth reagent adding. Each circular detection chamber 2 is also independent of each other, which is located at the outer circumferential position of the disc-shaped microfluidic chip. The main function of the circular detection chamber 2 is to allow the sample and various reagents to be fully mixed and reacted here to provide reliable reaction results for subsequent detection work.

[0048] The present embodiment also includes positioning holes and positioning pins, which are arranged at the inner ring position of the microfluidic chip in the form of a ring, and the positioning holes are arranged at the outer ring position. The positioning holes and the positioning pins are of great significance, which can realize accurate fixation and positioning of the microfluidic chip in the detection equipment and the sample adding equipment. Through this design, the position of the microfluidic chip in different equipment can be ensured to be accurate, thereby improving the accuracy and stability of detection and sample adding.

[0049] The V-shaped sample chamber 9 as the initial bearing area of the sample is directly connected with the horseshoe-shaped reagent chamber 4 through the first capillary valve 7. The first capillary valve 7 plays a key role in the whole chip fluid transmission process.

[0050] Similarly, the horseshoe-shaped reagent chamber 4 is directly connected with the circular detection chamber 2 through the second capillary valve 8. The design of the second capillary valve 8 is similar to that of the first capillary valve 7.

[0051] The present embodiment also includes an annular sealing film and a circular sealing film. The annular sealing film has a unique annular structure, and its main function is to seal the injection hole 6 or the air vent on the horseshoe-shaped reagent chamber 4 during the sample addition process. During the sample addition, the sealing effect of the annular sealing film can effectively prevent the leakage of the sample or the outside air, thereby ensuring the accuracy and safety of the sample addition process. The circular sealing film is used to seal the sample addition hole 10 and the air vent on the V-shaped sample chamber 9 during the reagent addition process. The circular sealing film can fit tightly against the sample addition hole 10 or the air vent, preventing the reagent from leaking during the addition process, while also avoiding the entry of external impurities, providing a good sealing environment for the entire detection process, and further ensuring the reliability of the test results.

[0052] From the perspective of ventilation structure, the disc-type microfluidic chip is provided with 16 first ventilation holes 1 and 16 second ventilation holes 3. Only the chip containing the sealing valve 5 has the second ventilation holes 3. These ventilation holes play a key role in the operation of the disc-type microfluidic chip. It can significantly increase the circulation of air above and below the disc-type microfluidic chip. In practical applications, disc-type microfluidic chips often need to perform rapid temperature change operations, and the good air circulation effect brought by the ventilation holes is particularly important. The smooth circulation of air can accelerate the transfer and exchange of heat, just like installing an efficient heat dissipation and heating system on the chip, so that the disc-type microfluidic chip can achieve temperature rise or fall more quickly, thereby improving the efficiency and accuracy of the entire detection or experimental process.

[0053] This embodiment also features a sealing valve 5, which performs its crucial sealing function after the sample loading operation is complete. It precisely seals the second capillary valve 8. During some reactions, aerosols may be generated. If these aerosols diffuse freely, they are highly likely to contaminate other components within the chip or the entire experimental environment, thereby impacting the reliability of the test results. The original design employed an iron wax valve. However, repeated testing revealed that during high-temperature amplification reactions, the paraffin wax within the valve melted, affecting not only the seal but also the reaction solution. Therefore, the valve was replaced with a sealing valve, sealed by injecting a UV-curing adhesive into the valve. This UV-curing adhesive should possess excellent biocompatibility, be non-cytotoxic, and not release substances that inhibit Taq enzyme activity or affect PCR reaction components such as the DNA template and primers. Furthermore, the physical properties of the adhesive should be compatible with the PCR reaction system; being too hard or too soft will not affect the stability of the reaction system, nor will it hinder the normal flow of the reaction solution. The hybrid sealing valve provided in this embodiment acts as a solid defense, effectively preventing the spread of aerosols and ensuring the purity of the reaction and the accuracy of the experimental results. However, the present invention also provides flexible options. If the user has other needs or is based on certain specific experimental conditions, the chip can be sealed using a second sealing method instead of a sealing valve. This flexible design fully considers the actual needs of different users and various complex experimental scenarios, making the disc-type microfluidic chip more widely applicable and more flexible in practical applications.

[0054] Embodiment 2: A sealing method for a disk-type microfluidic chip, the sealing method comprising the following steps:

[0055] Step 1: First, ensure the cleanliness and stability of the experimental environment, and prepare the required experimental equipment such as the sample diluent to be tested, pipette, etc. When the annular sealing film tightly seals the injection hole 6 on the horseshoe-shaped reagent chamber 4, this step is very critical. The sealed injection hole 6 can prevent external impurities from entering the horseshoe-shaped reagent chamber 4, ensuring the purity of subsequent reagents. At this time, pick up the pipette and carefully draw an appropriate amount of the prepared sample diluent to be tested. The operation of the pipette requires precise control to avoid drawing too much or too little sample diluent. Then, accurately align the tip of the pipette with the sample addition hole 10 on the V-shaped sample chamber 9, and slowly and steadily inject the sample diluent into the V-shaped sample chamber 9. During the injection process, closely observe the inflow of the sample diluent to ensure that it enters the V-shaped sample chamber 9 smoothly.

[0056] Step two, after the sample is added, the sample hole 10 on the V-shaped sample chamber 9 is sealed with the round sealing film. This operation can prevent the sample from leaking during the subsequent centrifugation process and ensure the integrity of the sample. After sealing, the disc microfluidic chip is placed in the centrifuge, and the centrifuge is started to apply centrifugal force. Under the action of centrifugal force, the sample diluent to be tested is like an invisible force pushing it to flow orderly along the first capillary valve 7 and the second capillary valve 8. The first capillary valve 7 and the second capillary valve 8 are designed to guide the sample diluent to flow smoothly from the V-shaped sample chamber 9 to the circular detection chamber 2. After a period of centrifugation, the sample diluent to be tested successfully enters the circular detection chamber 2, preparing for the subsequent reaction.

[0057] Step three, after the sample transfer is completed, the ring-shaped sealing film is carefully removed. During the removal process, attention should be paid to avoid unnecessary damage to the chip, and to prevent external impurities from entering the chip. After removing the ring-shaped sealing film, the pipette is picked up again, and the reagents required for the reaction are sequentially sucked according to the specific requirements of the experiment. The amount of each reagent should be strictly controlled according to the experimental scheme to ensure the accuracy of the reagent amount. Then, the tip of the pipette is inserted into the horseshoe-shaped reagent chamber 4 through the liquid injection hole 6, and the reagent is slowly injected. During the injection process, attention should be paid to observe the inflow of the reagent to ensure that the reagent is evenly distributed in the horseshoe-shaped reagent chamber 4.

[0058] Step four, after the reagent is added, the disc microfluidic chip is placed in the centrifuge again. Under the action of centrifugal force, the reagent required for the reaction is like a power, quickly moving through the second capillary valve 8 to the circular detection chamber 2. During the flow of the reagent in the second capillary valve 8, it is necessary to ensure that the channel is unobstructed to avoid blockage and other situations affecting the transfer of the reagent. After a period of centrifugation, the reaction reagent successfully enters the circular detection chamber 2 and mixes with the sample diluent to be tested previously, creating conditions for the reaction to proceed.

[0059] Step five, after the sample and the reagent enter the circular detection chamber 2, the reaction solution needs to be sealed to ensure that the reaction is carried out in a relatively closed environment.

[0060] The first sealing method is to seal the valve 5. Prepare a clean pipette and an appropriate adapter tip, and accurately draw the appropriate amount of UV-curable glue. Carefully insert the pipette tip into the sealing valve 5, slowly push the pipette plunger, and inject the UV-curable glue into the sealing valve 5. Observe the amount of glue during the injection process to avoid overflow due to excessive injection or incomplete filling of the second capillary valve 8 due to insufficient injection. After the injection is completed, quickly and smoothly place the chip in the centrifuge, and set the centrifugation time to 1 minute. Under the action of centrifugal force, the UV-curable glue in the sealing valve 5 begins to flow, gradually overcoming the surface tension and channel resistance, quickly and uniformly filling every corner of the second capillary valve 8, ensuring that there are no air bubbles remaining in the channel, and making the glue fully adhere to the channel wall, providing a good foundation for subsequent curing. After centrifugation, remove the chip. Hold the UV irradiation pen, accurately aim the pen end at the second capillary valve 8, and start from the beginning of the channel. Slowly and evenly irradiate the UV-curable glue in the channel. During the irradiation process, the specific wavelength light emitted by the UV irradiation pen is absorbed by the photoinitiator in the glue, triggering the polymerization reaction. As the irradiation continues, the originally liquid UV-curable glue gradually undergoes crosslinking reaction, changes from a flowing state to a solid state, and forms a solid and well-sealed glue structure, effectively sealing the second capillary valve 8. After sealing, place the chip on the table. During the placement process, ensure that the table is stable to avoid shaking or tilting the chip, which may affect the reaction. At this time, wait for the next step.

[0061] The second sealing method includes the following steps:

[0062] When the disc-type microfluidic chip is not provided with a sealing valve 5, an alternative sealing solution is required to ensure the integrity of the reaction environment. First, the previously removed annular sealing film is re-covered and accurately fitted on the injection hole 6 of the horseshoe-shaped reagent chamber 4, ensuring that the edge of the sealing film is completely aligned with the edge of the injection hole 6, and using the viscosity or pressure bonding method of the sealing film itself to form a physical barrier layer to prevent liquid from leaking from the injection hole 6 or external impurities from entering during the reaction. After completing the sealing of the injection hole 6 of the horseshoe-shaped reagent chamber 4, use a pipette to absorb the ultraviolet curing glue and slowly inject it through the sample addition hole 10 of the V-shaped sample chamber 9. During the injection process, the chamber state needs to be continuously observed until the ultraviolet curing glue completely fills the V-shaped sample chamber 9, forming a small protrusion higher than the plane of the sample addition hole 10. This operation not only ensures that the sample addition hole 10 is completely covered, but also uses excess colloid to form a more solid sealing layer. After the injection is completed, turn on the ultraviolet lamp located in the center of the chip for 60 seconds of irradiation curing. During these 60 seconds, the photosensitive initiator in the UV-curing glue absorbs ultraviolet energy, triggering a polymerization reaction, and the colloid gradually changes from liquid to solid, eventually forming a hard, dense sealing layer on the surface of the sample addition hole 10, which effectively isolates the external environment from the material exchange inside the chip. At the same time, the horseshoe-shaped reagent chamber 4 injection hole 6 will be reinforced twice using traditional mechanical sealing methods during the entire reaction detection process. This mechanical seal is usually composed of components such as a clamp and a sealing ring. The sealing ring is squeezed by external force to fit tightly around the injection hole 6, further enhancing the sealing effect. The advantage of mechanical sealing is that it can be adjusted and disassembled multiple times according to actual needs, providing convenience when reagents need to be replenished or other operations need to be performed. After the UV-curing glue is completely cured and the mechanical seal is installed and debugged, hold the two sides of the chip lightly with both hands, keep it horizontal, and carefully place the chip flat on a clean, stable table. Avoid violent vibration or tilting during placement to prevent cracks or liquid displacement in the sealed colloid. At this point, the chip has completed all sealing operations and can wait for the next reaction process to start, ensuring that the entire experimental process is carried out in a stable and closed environment, thereby ensuring the accuracy and reliability of the test results.

[0063] Those skilled in the art will understand that the above description is only a preferred embodiment of the present invention, and the features described in the various embodiments and / or technical solutions of the present disclosure can be combined or combined in various ways, even if such a combination or combination is not explicitly described in the present disclosure. It is not intended to limit the present invention. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art can still modify the technical solutions described in the aforementioned embodiments, or make equivalent substitutions for some of the technical features therein. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

[0064] While the preferred embodiments of the application have been described, additional variations and modifications can be made to these embodiments by those skilled in the art once they have the benefit of the foregoing description without departing from the spirit and scope of the application. Accordingly, the appended claims are intended to encompass all variations and modifications made to the embodiments within the scope of the application. It is apparent that those skilled in the art can modify and adapt the application in various ways without departing from the spirit and scope of the application. It is therefore intended that the application be construed in all its aspects as including all such modifications and variations as fall within the scope of the present application and equivalents thereof.

Claims

1. A disk-type microfluidic chip, characterized in that: The disc-type microfluidic chip includes 16 reaction detection modules, which are evenly distributed on the circumference of the disc-type microfluidic chip. Each reaction detection module includes a chamber and a channel, specifically including a first ventilation hole (1), a circular detection chamber (2), a second ventilation hole (3), a horseshoe-shaped reagent chamber (4), a sealing valve (5), an injection hole (6), a first capillary valve (7), a second capillary valve (8), a V-shaped sample chamber (9), and a sample addition hole (10); Each V-shaped sample chamber (9) is connected to each other. The V-shaped sample chamber (9) is arranged on one side of the disc-type microfluidic chip close to the center of the circle. The terminal positions on both sides of the V-shaped sample chamber (9) are respectively provided with a sample addition hole (10) or a vent hole. If the terminal on one side is a sample addition hole (10), the terminal on the other side is regarded as a vent hole. The horseshoe-shaped reagent chamber (4) is located in the middle of the disc-type microfluidic chip, and each horseshoe-shaped reagent chamber (4) is provided with two injection holes (6) or vents, wherein one end is the injection hole (6) and the other end is regarded as the vent; The circular detection chamber (2) is located on the periphery of the disc-type microfluidic chip and is used to mix the sample and the reagent and generate a reaction; The V-shaped sample chamber (9) is connected to the horseshoe-shaped reagent chamber (4) via a first capillary valve (7); The horseshoe-shaped reagent chamber (4) is connected to the circular detection chamber (2) via a second capillary valve (8); The disc-type microfluidic chip further comprises a sealing valve (5), and ultraviolet curing glue is injected into the sealing valve (5) to seal the second capillary valve (8).

2. The disk-type microfluidic chip according to claim 1, characterized in that: The disc-shaped microfluidic chip further comprises positioning holes and positioning pins. The inner ring of the disc-shaped microfluidic chip is provided with positioning pins, and the outer ring is provided with positioning holes.

3. The disk-type microfluidic chip according to claim 1, characterized in that: The disc-type microfluidic chip further comprises an annular sealing film and a circular sealing film. The annular sealing film is used to seal the sample addition hole (10) on the horseshoe-shaped reagent chamber (4), and the circular sealing film is used to seal the two injection holes (6) on the V-shaped sample chamber (9).

4. The disk-type microfluidic chip according to claim 1, characterized in that: The disc-type microfluidic chip is provided with 16 first ventilation holes (1) and 16 second ventilation holes (3), and only the disc-type microfluidic chip containing the sealing valve (5) has the second ventilation holes (3).

5. A sealing method for a disk-type microfluidic chip, characterized in that: The method is implemented based on the disk-type microfluidic chip according to any one of claims 1 to 4, and comprises the following steps: S1. With the annular sealing film sealing the injection hole (6) on the horseshoe-shaped reagent chamber (4), align the tip of the pipette with the sample injection hole (10) on the V-shaped sample chamber (9) and inject the sample diluent into the V-shaped sample chamber (9); S2. Use a circular sealing film to seal the sample loading hole (10) on the V-shaped sample chamber (9). After the sealing is completed, place the disc-type microfluidic chip in a centrifuge, turn on the centrifuge to apply centrifugal force, and the sample dilution to be tested successfully enters the circular detection chamber (2); S3. After the sample transfer is completed, the annular sealing film is removed. After removing the annular sealing film, the pipette is picked up again, and the reagents required for the reaction are drawn in sequence. The tip of the pipette is inserted into the horseshoe-shaped reagent chamber (4) through the injection hole (6), and the reagents are added to the horseshoe-shaped reagent chamber (4); S4. After the reagents are added, the disc-shaped microfluidic chip is placed in a centrifuge and moved to the circular detection chamber (2) through the second capillary valve (8). The reaction reagent enters the circular detection chamber (2) and mixes with the dilution solution of the sample to be tested that enters in step 3. S5. After the sample and reagent enter the circular detection chamber (2), the reaction solution is sealed.

6. The sealing method of the disk-type microfluidic chip according to claim 5, characterized in that: The sealing of the reaction solution in S5 includes a sealing valve sealing method and a physical barrier layer method, wherein the sealing valve sealing method is: S1.

1. Prepare a clean pipette and an adapter tip, absorb an appropriate amount of UV curing glue, insert the pipette tip into the sealing valve (5), push the pipette push rod, and inject the UV curing glue into the sealing valve (5); after the injection is completed, place the chip steadily in a centrifuge and set the centrifugation time to 1 minute to quickly and evenly fill the second capillary valve (8); S1.

2. After the centrifugation is completed, the disc-type microfluidic chip is taken out, and the tip of the UV irradiation pen is held and aimed at the second capillary valve (8). Starting from the starting end of the second capillary valve (8), the UV curing glue in the second capillary valve (8) is irradiated in sequence at a uniform speed; S1.

3. As the irradiation continues, the originally liquid UV curing glue gradually undergoes a cross-linking reaction and gradually changes from a fluid state to a solid state, forming a strong colloidal structure with good sealing performance, thereby sealing the second capillary valve (8). After the sealing is completed, the disc-type microfluidic chip should be taken out of the centrifuge and placed flat on the table.

7. The sealing method of a disk-type microfluidic chip according to claim 6, characterized in that: The physical barrier layer method comprises the following steps: S2.

1. Re-cover the removed annular sealing film and attach it to the injection hole (6) of the horseshoe-shaped reagent chamber (4), using the adhesiveness of the sealing film itself or pressure bonding to form a physical barrier layer; S2.

2. After completing the sealing of the injection hole (6) of the horseshoe-shaped reagent chamber (4), use a pipette to absorb the UV-curing glue and slowly inject it through the sample injection hole (10) of the V-shaped sample chamber (9). During the injection process, the chamber state needs to be continuously observed until the UV-curing glue completely fills the V-shaped sample chamber (9) to form a bulge higher than the plane of the sample injection hole (10); S2.

3. After the injection is completed, turn on the UV lamp located in the center of the chip for 60 seconds of irradiation curing. The colloid gradually changes from liquid to solid, and finally forms a sealing layer on the surface of the sample hole (10).

Images