A microfluidic chip

Abstract

The present application relates to a kind of microfluidic chip, comprising: chip body, including flat reaction zone, the reaction zone includes chip import and chip export, long and narrow passage is included between the reaction zone and the chip export;Housing body, including for accommodating soluble microsphere housing cavity, the housing cavity includes first import, second import and first export, the first import is used to receive sample, the first export is connected with the chip import of the reaction zone;Piston is configured to push the sample after the microsphere in the housing cavity is dissolved into the reaction zone by the second import;And deformable material is sealed after deformation the chip import of the reaction zone.The microfluidic chip of the present application can make the sample after microsphere dissolution into the reaction zone by different sealing means, and the chip import and chip export of reaction zone are sealed, form full-closed system, can reduce interference.

Description

Technical Field

[0001] This invention relates to the field of biomedical detection technology, and in particular to a microfluidic chip. Background Technology

[0002] Traditional PCR technology suffers from numerous drawbacks, including high equipment costs and demanding laboratory requirements. Microfluidic technology, based on microfabrication techniques, enables micro-volume reactions, significantly reducing detection costs and greatly improving detection efficiency. However, existing microfluidic chips still suffer from problems such as poor sealing, susceptibility to interference, heavy reliance on external pressure devices, and cumbersome operation. Summary of the Invention

[0003] To address the technical problems existing in the prior art, this invention proposes a microfluidic chip, comprising: a chip body including a flat reaction region, the reaction region including a chip inlet and a chip outlet, and an elongated channel between the reaction region and the chip outlet; a container including a cavity for accommodating soluble microspheres, the cavity including a first inlet, a second inlet and a first outlet, the first inlet being used to receive a sample, and the first outlet being connected to the chip inlet of the reaction region; a piston configured to push a sample dissolved from the microspheres in the cavity into the reaction region through the second inlet; and a deformable material that, after deformation, seals the chip inlet of the reaction region.

[0004] The microfluidic chip described above has a reaction zone with a depth of 200-450 μm and a volume of 10-15 μl; the thermal conductivity of at least a portion of the chip body material is such that, at a heater temperature of 95°C, the average heating rate of the reaction zone is ≥15°C / second.

[0005] In the microfluidic chip described above, the cross-sectional area of ​​the elongated channel is 0.07-0.2 mm². 2 The length is 15-25mm.

[0006] As described above, in the microfluidic chip, the accommodator includes a first channel connected to the first inlet; a second channel connected to the second inlet, the second channel being adapted to the piston; the first channel being connected to the second channel; and the piston closing the first channel after passing through the first channel.

[0007] The microfluidic chip described above, wherein the accommodator includes a removable injection cap that is adapted to and closes the first inlet.

[0008] In the microfluidic chip described above, when the piston pushes the dissolved sample of the microspheres in the accommodating cavity into the reaction zone, the deformable material gradually deforms.

[0009] In the microfluidic chip described above, when the piston pushes the sample after all the dissolved microspheres in the accommodating cavity into the reaction zone, the deformable material has not yet sealed the chip inlet of the chip body.

[0010] In the microfluidic chip described above, when the piston pushes the sample after all the dissolved microspheres in the accommodating cavity into the reaction zone and continues to move, the deformable material continues to deform and seal the chip inlet.

[0011] The microfluidic chip described above, wherein the accommodator includes an exhaust channel connected to the chip outlet, and the exhaust channel is provided with a water-sensitive seal.

[0012] In the microfluidic chip described above, after the piston pushes the sample containing all the dissolved microspheres in the accommodating cavity into the reaction zone, the deformable material seals the chip outlet after deformation.

[0013] The microfluidic chip described above further includes a sealing plug, wherein, after the piston pushes the sample containing all the dissolved microspheres in the accommodating cavity into the reaction zone, the sealing plug moves and seals the chip outlet.

[0014] In the microfluidic chip described above, the direction of movement of the piston is parallel to the direction of the chip body.

[0015] The microfluidic chip described above, wherein the accommodator includes a chip interface for accommodating the chip body, the chip body and the chip interface are interference-fitted, and the deformable material is disposed within the chip interface.

[0016] In the microfluidic chip described above, the direction of movement of the piston is perpendicular to the direction of the chip body.

[0017] The microfluidic chip as described above, wherein the chip body is attached to the accommodating body, the chip inlet of the chip body corresponds to the first outlet of the accommodating cavity, and the deformable material is disposed between the chip inlet and the first outlet.

[0018] The microfluidic chip described above further includes an exhaust sealing piston, wherein the deformable material includes an exhaust port communicating with the chip outlet. When the piston pushes the sample after all the dissolved microspheres in the accommodating cavity into the reaction zone and the deformable material seals the chip inlet after deformation, the exhaust sealing piston seals the exhaust port of the deformable material.

[0019] In the microfluidic chip described above, the vent of the deformable material includes a protruding lip with a notch for venting, and the vent sealing piston enters the lip to disable the notch.

[0020] The microfluidic chip described above further includes one or more additional pistons, the chip body includes one or more additional flat reaction zones, the accommodator includes one or more additional accommodating cavities, and the one or more additional pistons are configured to push a sample of dissolved microspheres from the corresponding one or more additional accommodating cavities into the corresponding one or more additional reaction zones.

[0021] The microfluidic chip described above, wherein the sample is a sample that has undergone nucleic acid extraction, and the sample is dissolved in the microspheres to obtain a system suitable for PCR reaction.

[0022] The microfluidic chip described above, wherein the PCR reaction system after the microspheres are dissolved is used for HPV detection.

[0023] According to another aspect of this application, a sealing device for use with a microfluidic chip as described above is proposed, comprising: a first clamp including: a first seat; and a first body and a second body located on the first seat; wherein a slit is included between the first body and the second body, the slit being adapted to receive the chip body of the microfluidic chip, and the first body and the second body being adapted to receive the receiving body of the microfluidic chip; a second clamp including: a second seat; a first pillar and a second pillar located on the second seat; a support located on the second seat; and a pressure block connected to the support; wherein the space between the first pillar and the second pillar is adapted to receive the first clamp; the pressure block is configured to move relative to the support, thereby pushing the piston of the microfluidic chip.

[0024] In the sealing device described above, the height between the first seat and the first body and the second body is a first height, and the height between the chip body of the microfluidic chip and the edge of the accommodating body that is supported on the first body and the second body is a second height. The first height is greater than the second height, and the height difference between the first height and the second height is equivalent to the deformation of the deformable material.

[0025] According to another aspect of this application, a method for operating a microfluidic chip as described above is proposed, comprising: adding a sample after nucleic acid extraction from the first inlet of the container of the microfluidic chip, wherein the sample after nucleic acid extraction dissolves microspheres in the container cavity to form a reaction system suitable for PCR; pushing the piston of the microfluidic chip to push the sample after dissolving the microspheres into the reaction zone of the microfluidic chip; and transferring the microfluidic chip into a PCR instrument.

[0026] The method described above further includes: after pushing the piston of the microfluidic chip to push the sample after dissolving the microspheres into the reaction zone of the microfluidic chip, further pushing the piston to seal the chip inlet of the reaction zone.

[0027] In the method described above, the detection time of the PCR instrument is within 2 hours, within 1.5 hours, or within 1 hour.

[0028] The microfluidic chip of this application can seal the chip inlet and outlet of the reaction zone after the sample dissolved by the microspheres enters the reaction zone through different sealing methods, forming a fully enclosed system. This can prevent external impurities from affecting the PCR reaction, improve the specificity and accuracy of detection, and largely avoid the dependence of existing microfluidic chips on external devices, thereby expanding the application scope of the microfluidic chip of this invention. Attached Figure Description

[0029] The preferred embodiments of the present invention will now be described in further detail with reference to the accompanying drawings, wherein: Figures 1A-1C This is a schematic diagram of a microfluidic chip according to an embodiment of this application; Figure 2 For the purposes of this application Figures 1A-1C Exploded view of the microfluidic chip in the embodiment; Figure 3A and Figure 3B This is a schematic diagram of a chip body structure according to an embodiment of this application; Figures 4A-4C This is a schematic diagram of a receiving body structure according to an embodiment of this application; Figure 5 This is a schematic diagram of a piston structure according to an embodiment of this application; Figure 6 This is a schematic diagram of the sample inlet cap structure according to an embodiment of this application; Figure 7 This is a schematic diagram of a piston limiting block structure according to an embodiment of this application; Figures 8A-8H This is a schematic diagram of different states of a microfluidic chip according to an embodiment of this application; Figure 9 This is a schematic diagram of a sealing device structure according to an embodiment of this application; Figure 10A and Figure 10B This is a schematic diagram of a microfluidic chip according to another embodiment of this application; Figure 11 for Figure 10A and Figure 10B Exploded view of the microfluidic chip in the embodiment; Figure 12 This is a schematic diagram of a deformable material structure according to another embodiment of this application; Figures 13A-13F This is a schematic diagram of different states of a microfluidic chip according to another embodiment of this application; Figure 14A and Figure 14B This is a schematic diagram of a microfluidic chip according to another embodiment of this application; Figure 15 for Figure 14A and Figure 14B Exploded view of the microfluidic chip in the embodiment; Figures 16A-16D A schematic diagram of different states of a microfluidic chip according to another embodiment of this application; and Figure 17 This is a schematic diagram of the operation process of a microfluidic chip according to an embodiment of this application. Detailed Implementation

[0030] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0031] In the following detailed description, reference can be made to the accompanying drawings, which form part of this application and illustrate specific embodiments of the present application. In the drawings, similar reference numerals describe substantially similar components in different figures. Specific embodiments of the present application are described in sufficient detail below to enable those skilled in the art to implement the technical solutions of the present application. It should be understood that other embodiments may also be utilized, or structural, logical, or electrical changes may be made to the embodiments of the present application.

[0032] The following disclosure provides many different embodiments or examples for implementing various features of this application. Specific examples of components and configurations are described below to simplify the disclosure. Of course, these are merely examples and are not intended to limit the scope of this application. For example, the following description of forming a first feature on or above a second feature may include embodiments where the first and second features are in direct contact, or embodiments where other features are formed between the first and second features, thus the first and second features are not in direct contact. Furthermore, component symbols and / or letters may be repeated in different examples within this application. This repetition is for simplification and clarity, and does not govern the relationship between the different embodiments and / or the architectures discussed.

[0033] Furthermore, this application may use spatial correspondence terms, such as "below," "lower," "lower," "higher," "higher," and similar terms, to describe the relationship between one component or feature and another component or feature in the drawings. Spatial correspondence terms are used to include different orientations of the device in use or operation, in addition to those described in the drawings. The device may be positioned (rotated 90 degrees or otherwise), and the spatial correspondence descriptions used in this application may be interpreted accordingly.

[0034] Although the numerical ranges and parameters disclosed in this broad disclosure are approximate, the values ​​described in the specific embodiments are as precise as possible. However, any numerical value inherently includes some error due to the standard deviation obtained from individual test measurements. Furthermore, as herein, "about" generally means within 10%, 5%, 1%, or 0.5% of a given value or range. Alternatively, the term "about" means within an acceptable standard error of an average value considered by those generally skilled in the art. Except in operational / working examples, or unless specifically indicated, all numerical ranges, quantities, values, and proportions disclosed herein, such as amounts of material, time periods, temperatures, operating conditions, proportions of quantities, and the like, should be understood to be modified by the term "about" in all cases. Therefore, unless stated to the contrary, the numerical parameters of this disclosure and the claims are approximate values ​​that may vary as needed. Each numerical parameter should be interpreted at least according to the number of significant figures reported and with the application of common rounding techniques. In this document, a range may be expressed as from one endpoint to another, or between two endpoints. Unless otherwise stated, all ranges disclosed herein include endpoints.

[0035] Figures 1A-1C This is a schematic diagram of a microfluidic chip according to an embodiment of this application. Figure 2 For the purposes of this application Figures 1A-1CAn exploded view of the microfluidic chip described in this embodiment. As shown, the microfluidic chip 100 includes a chip body 110, a container 120, multiple pistons 131-133, and a deformable material 140. The container 120 includes multiple cavities 121-123. The chip body 110 includes multiple reaction zones 111-113. Each cavity includes a first inlet for receiving a sample, a second inlet for the piston to enter, and a first outlet communicating with the reaction zone in the chip body. Each cavity contains soluble microspheres. When a sample enters the cavity, the microspheres can rapidly collapse and dissolve, forming a reaction system suitable for PCR. The piston enters the cavity through the second inlet and further pushes the sample after microsphere dissolution into the reaction zone corresponding to that cavity. When or after the sample after microsphere dissolution enters the reaction zone, the deformable material 140 deforms, sealing the chip inlet of the reaction zone.

[0036] In some embodiments, the microspheres include specific primer pairs (Forward & Reverse) for a specific target, fluorescent probes (e.g., TaqMan probes, Molecular Beacons probes, FRET Hybridization probes, Scorpions Probes, Sunrise / Eclipse Probes, LNA / MB modified probes, etc.), and a premixed PCR reaction system (e.g., containing dNTPs, essential enzymes, Mg²⁺, etc.). In some embodiments, the microspheres may include one or more other auxiliaries, such as protectants, preservatives, etc. In some embodiments, the microspheres are one or more spheres with a large diameter, or granules or powders. In some embodiments, the microspheres can be formed by various methods such as freeze-drying, spray drying, supercritical fluid drying, high-pressure pulsed fluidized bed granulation, etc. As understood, the microfluidic chip of the present invention can be used for PCR reactions of various types and systems; the above is merely an illustrative example and does not limit the scope of the invention. For those skilled in the art, based on the desired PCR or qPCR reaction, a PCR reaction system can be constructed in the manner of this invention, and microspheres suitable for the microfluidic chip of this invention can be manufactured and obtained, thereby being applied in this invention.

[0037] Furthermore, unlike ordinary microfluidic chips, the microfluidic chip of this invention receives samples that have undergone nucleic acid extraction. Therefore, the microfluidic chip of this invention does not include functions or facilities related to nucleic acid extraction. This design greatly simplifies the structure of the microfluidic chip, offering advantages in reducing complexity and improving reliability for sealing control and operation. Moreover, existing nucleic acid extraction operations are already widespread and will not affect the application of this invention. It is also worth noting that the sample received by this invention forms a PCR reaction system immediately after dissolving the microspheres, without sample dilution, which is detrimental to reducing non-specific signal levels. Specific experimental results demonstrate the feasibility of the scheme for the microfluidic chip of this invention, and the sensitivity of the detection results can reach a very high level.

[0038] In this embodiment, the deformable material 140 is used to seal the chip inlet of the chip body 110. In some embodiments, the deformable material 140 may be disposed between the first outlet of the accommodating body 120 and the chip inlet of the chip body 110 and includes a hole connecting the first outlet of the accommodating body 120 and the chip inlet of the chip body 110. When the deformable material 140 is under pressure, it deforms, compressing the hole until it completely disappears, thereby achieving a seal. In other embodiments, the piston may directly abut against the hole connecting the first outlet of the accommodating body 120 and the chip inlet of the chip body 110. Since the diameter of the piston is larger than the diameter of the hole, when pressure is applied by the piston, the deformable material 140 between the piston and the chip inlet deforms, thereby sealing the gap between the piston and the chip inlet. In some embodiments, the deformable material 140 may be silicone (PDMS), butyl rubber (IIR), thermoplastic elastomer (e.g., TPU or SEBS), fluororubber and perfluorinated elastomer (e.g., FFKM or FEP / PFA), or cyclic olefin copolymer / cyclic olefin polymer (COC / COP).

[0039] In this embodiment, the piston moves in a direction parallel to the direction of the chip body. The chip body 110 is disposed on the first side of the container 120; pistons 131-133 are disposed on the second side of the container 120. The first side and the second side are opposite sides. Pistons 131-133 move from the first side of the container 120 into the container 120, pushing the sample after the microspheres have dissolved into the chip body 110 on the second side. A deformable material 140 is disposed between the chip body 110 and the container 120. When the sample after the microspheres have dissolved is pushed into the chip body 110, the deformation of the deformable material seals the chip body and the container. In this embodiment, the arrangement of the piston and the chip body on opposite sides of the container means that the sample does not need to change direction when flowing into the reaction area of ​​the chip body, which is more conducive to achieving a seal and also more conducive to the integration of various structural parts of the microfluidic chip.

[0040] In this embodiment, the microfluidic chip includes three sets of detection units. Each detection unit includes a piston, a cavity in a corresponding container, and a corresponding reaction zone. Therefore, the microfluidic chip of this embodiment can simultaneously detect three different samples, improving detection efficiency. As understood, the present invention does not limit the number of detection units: there can be one, two, three, or more detection units.

[0041] In some other embodiments, the same cavity may correspond to multiple reaction zones (i.e., the cavity includes at least one second outlet connected to another reaction zone), allowing the same PCR reaction to be performed in multiple reaction zones, thereby making the detection results more reliable. In some other embodiments, the same cavity may also correspond to multiple pistons, resulting in more uniform pressure within the cavity and preventing improper operation.

[0042] In some embodiments, although the piston-pushing operation in the microfluidic chip of the present invention can be performed by a dedicated device, avoiding uneven force control during manual operation, it can still be performed manually under limited conditions. Even with dedicated devices, as described later in this application, the structure of the dedicated devices is relatively simple. This largely avoids the dependence of existing microfluidic chips on external devices, thereby expanding the application scope of the microfluidic chip of the present invention.

[0043] In some embodiments, after the sample dissolved by the microspheres is pushed into the reaction zone and sealed, the microfluidic chip of the present invention can be transferred to a real-time PCR instrument for PCR reaction and fluorescence detection, and the detection results can be output.

[0044] Figure 3A and Figure 3B This is a schematic diagram of a chip body structure according to an embodiment of this application. (In conjunction with...) Figure 3A and Figure 3B The chip body 110 may include multiple reaction zones 111-113, which can be used to contain samples dissolved by the microspheres and to perform PCR reactions. In some embodiments, the reaction zones 111-113 are generally flat, with a depth of approximately 200-450 μm and a capacity of approximately 10-15 μl. The flat shape facilitates heat transfer in the reaction zones, allowing for rapid temperature increases or decreases, thereby shortening the detection time.

[0045] In some embodiments, taking reaction zone 111 as an example, it includes a chip inlet 114, a chip outlet 115, and a residence zone 116. The chip inlet 112 is connected to the first end of the residence zone 116, which is the main area for containing the sample after the microspheres have dissolved. The chip outlet 113 is connected to the second end of the residence zone 116 via a narrow channel 117. When the sample after the microspheres have dissolved enters the residence zone 116, the gas in the residence zone 116 is discharged through the narrow channel 117 and then through the chip outlet 115. In some embodiments, the chip inlet 112 and the residence zone 116 can also be connected via a narrow channel. In some embodiments, the length of the narrow channel between the chip inlet and the residence zone 116 can be greater than the length of the narrow channel between the chip outlet 113 and the residence zone 116. In some embodiments, the size of the narrow channel can be on the micrometer scale, and its cross-sectional area can be 0.07-0.2 μm. 2 The length can be 15-25 μm. The elongated channel's size serves two main purposes: firstly, it acts as a "capillary brake," preventing high-speed liquid ejection or backflow that could cause contamination; secondly, the elongated channel stabilizes pressure, facilitating the sealing of the chip outlet 115; and thirdly, it retains most of the sample in the residence area 116, which is beneficial for the PCR reaction. In some embodiments, a water-reactive sealing material is provided at the chip outlet 115. When the sample reaches the water-reactive sealing material through the elongated channel, the material changes, sealing the chip outlet 115. In some embodiments, the water-reactive sealing material includes an expanded PTFE (ePTFE) waterproof and breathable membrane.

[0046] As those skilled in the art understand, PCR reactions mainly involve multiple cycles of three reaction steps: deformation, annealing, and extension, achieving in vitro amplification of gene samples through temperature control. To improve detection speed, rapidly increasing or decreasing the temperature of the microfluidic chip reaction zone is highly advantageous. Therefore, in some embodiments, the chip body 110 is made of a material with high thermal conductivity to meet the rapid temperature changes required in accelerating the PCR reaction. In some embodiments, the high thermal conductivity of the chip body 110 material ensures that, with an external heater temperature of 95°C, the average heating rate of the reaction zone is ≥15°C / second, with a maximum heating rate ≥30°C / second; and with an external heater temperature of 10°C, the maximum cooling rate of the reaction zone is ≥10°C / second.

[0047] Meanwhile, the material of the chip body 110 must also meet the requirements of microfabrication, enabling the reaction area to be easily fabricated on it. Furthermore, to avoid affecting fluorescence detection, the material of the chip body 110 should also have high light transmittance. In some embodiments, the high thermal conductivity material of the chip body 110 includes silicon wafers (high-purity silicon), silicon nitride, aluminum nitride, or silicon carbide, which can rapidly and uniformly transfer the heat required for PCR amplification, reducing the conventional PCR reaction time from several hours to about 30 minutes, and also supporting micron-level channel etching.

[0048] In some embodiments, the chip body 110 may include a main material 118 and a cover 119. The main material 118 may be made of materials such as silicon wafers (high-purity silicon), and structures such as reaction regions and elongated channels can be easily fabricated on the main material 118 using wafer processing techniques (e.g., etching), ensuring smooth and burr-free flow channels and thus avoiding sample residue. The cover 119 is made of glass with a light transmittance ≥90% to facilitate multicolor fluorescence signal acquisition. The glass is bonded to the main material 119 through a high-temperature anodic bonding process (temperature 300-400℃, voltage 500-1000V) to achieve a sealed bond between the two, forming a closed flow channel system. The bonding strength between the two is ≥15MPa, which can prevent the chip body from delaminating at high temperatures. In some embodiments, the surface of the main material 118 may also be coated with a hydrophilic film with a hydrophilic angle of less than 30 degrees, ensuring that the sample after the microspheres are dissolved can smoothly enter the residence area within the elongated channel. In some embodiments, the dimensional tolerance of the chip body is ≤0.03mm and the flatness is less than 5um, which can effectively reduce assembly errors.

[0049] Figures 4A-4C This is a schematic diagram of a container structure according to an embodiment of this application. Referring to Figure 4, the container 120 includes multiple cavities 121-123, which can be used to contain microspheres and form a PCR reaction system. In this embodiment, each cavity contains a single soluble microsphere produced by a freeze-drying process. When the microspheres come into contact with a sample extracted with nucleic acid, they can rapidly collapse and dissolve to form a PCR reaction system that can be used for direct amplification.

[0050] In some embodiments, taking the accommodating cavity 121 as an example, it includes a first inlet 1211, a second inlet 1212, and a first outlet 1213. The first inlet 1212 is used to receive samples, the second inlet 1211 is used to allow the piston to enter, and the first outlet 1213 is used to communicate with the chip inlet of the reaction zone. In some embodiments, the second inlet 1212 is connected to the accommodating cavity 121 through a second channel 1214, while the first inlet 1211 is connected to the second channel 1214 through a first channel 1215, so that the nucleic acid extracted sample can enter the second channel 1214 through the first inlet 1211 and then enter the accommodating cavity 121 through the second channel 1215. The second channel 1214 is adapted to the size of the piston, and the piston can move along the second channel 1214 and enter the accommodating cavity 121. In some embodiments, when the piston enters the accommodating cavity 121 along the second channel 1214, it first passes through the first channel 1215 and thereby closes the first channel 1215, thus effectively preventing external substances from entering the accommodating cavity through the first inlet.

[0051] In some embodiments, the container 120 includes an outlet channel disposed between the container cavity 121 and the first outlet 1213. The outlet channel ensures that the sample in the container cavity 121 can be fully inserted into the reaction zone of the chip body under the push of the piston. In some embodiments, the piston tip includes a protrusion that matches the outlet channel and can enter the outlet channel to assist in sealing the outlet channel. Furthermore, since the compression of the deformable material is performed integrally by the container 120 and the chip body 110, a larger deformation can be generated, which is beneficial to the sealing of the chip inlet of the chip body 110.

[0052] In some embodiments, the accommodator 120 includes a chip interface 124 for accommodating the chip body 110. One end of the chip body 110, where a chip inlet and a chip outlet are located, is accommodated in the chip interface 124. In some examples, the chip interface is a recess, into which the chip body is inserted, and the two are connected and sealed by an interference fit (interference amount 0.01-0.05 mm) without any adhesive, and will not loosen under no external force. This ensures a seal without causing contamination. The deformable material 140 is a perforated silicone pad disposed in the chip interface 124. When compressed, the silicone pad deforms, filling the space between the chip interface 124 and the chip body 110, thus sealing the chip inlet of the reaction zone.

[0053] In some embodiments, the container 120 further includes multiple exhaust channels 125-127, which correspond to reaction zones 111-113 respectively, connecting the chip outlet of each reaction zone to the outside for discharging gas from the reaction zone. In some embodiments, the container 120 may also include a water-reactive seal 128, which may be disposed in or at the end of the exhaust channel. When the water-reactive seal is not in contact with water, only gas is allowed to pass through; when the water-reactive seal comes into contact with water, it seals the exhaust channel, effectively preventing the sample in the reaction zone from leaving the exhaust channel and contaminating the external environment. In some embodiments, the water-reactive seal may be a porous sintered polyethylene (PE) self-sealing filter element, which uses a special process to embed a polymer elastomer into the porous structure of the PE sintered filter element. In a dry state, the polymer elastomer does not expand, and the air passages between the PE networks maintain high air permeability; after contact with moisture, the polymer elastomer expands rapidly, blocking the porous structure, preventing aqueous solutions or gases from passing through, and sealing the exhaust channel.

[0054] In some embodiments, the accommodating body 120 may further include a set of engaging portions 129, which cooperate with another set of engaging portions 138 on the piston 130. When the piston moves along the second channel 1214 into the accommodating cavity 121, the two sets of engaging portions engage with each other to fix the position of the piston. In the case of multiple pistons, the two sets of engaging portions can simultaneously fix the positions of multiple pistons. In some embodiments, the material of the accommodating body may be high-temperature resistant engineering plastics such as PC, PP, and POM.

[0055] Figure 5 This is a schematic diagram of a piston structure according to an embodiment of this application. (In conjunction with...) Figure 5 Multiple pistons 131-133 form a set of pistons. A pusher plate 134 is disposed on the pistons 131-133 to fix the set of pistons together, so that pushing the pusher plate 134 can simultaneously push the pistons 131-133 into the container 120, thereby simultaneously pushing multiple samples of dissolved microspheres in multiple accommodating cavities into multiple reaction zones. In some embodiments, a single piston can also be pushed independently to provide a more flexible operating mode.

[0056] In some embodiments, taking piston 131 as an example, it includes a plunger 135 and a piston head 136. The plunger 135 is housed in a second channel 1214 connected to the receiving cavity 121, and pushing the pusher plate 134 allows the plunger 135 to move within the second channel 1214. The size of the plunger 135 is approximately equal to the size of the second channel 1214, allowing the plunger 135 to slide with damping along the second channel 1214, closing the first channel 1215 when passing through it for receiving samples. In some embodiments, the piston head 136 is located at the end of the plunger away from the pusher plate, and is used to abut against the bottom of the receiving cavity 121, assisting in closing the first outlet of the receiving cavity 121. In some embodiments, the end of the piston head 136 away from the plunger includes an outwardly extending protrusion 137, which can close the first outlet of the receiving cavity after the piston pushes the pre-reaction liquid into the chip body, thereby sealing the outlet channel. In some embodiments, the protrusion 137 may engage with the deformable material 140 to seal the chip inlet of the chip body.

[0057] In some embodiments, the deformable material 140 includes two sets of openings 141-143 and 144-146, which correspond to the positions of the chip inlet and chip outlet of each reaction zone, respectively, allowing the chip inlet and chip outlet to communicate with the outlet and exhaust channel of the accommodating cavity, respectively. During the piston insertion into the accommodating cavity, the deformable material 140 is slightly pressurized and undergoes partial deformation, gradually reducing the size of the openings. After the piston is inserted into the accommodating cavity, the deformable material 140 continues to be pressurized, undergoing greater deformation, filling the space between the accommodating chip interface and the chip body's entry chip interface end, causing all openings to disappear, thereby sealing the chip inlet and / or chip outlet and preventing sample leakage during PCR amplification. In some embodiments, the deformable material can be a silicone sealing strip. In some embodiments, the thickness of the silicone sealing strip can be 0.5-1.0 mm.

[0058] In some embodiments, to prevent external contaminants from entering the accommodating cavity through the first channel before sample addition, the microfluidic chip may also include a removable injection cap 150. Before sample addition, the injection cap 150 covers and seals the first inlet of the accommodating cavity. When sample addition is required, the injection cap can be removed. In some embodiments, the injection cap 150 includes a plurality of sealing posts 151-153, the size of which is comparable to the size of the first channel. When the injection cap 150 is placed on the microfluidic chip, the plurality of sealing posts 151-153 respectively enter the plurality of first channels and seal them.

[0059] Figure 6 This is a schematic diagram of a sample inlet cap structure according to one embodiment of this application. (In conjunction with...) Figure 6The injection cap 150 includes a cover plate 154, which can cover multiple first inlets of the container to seal the first inlets. In some embodiments, the injection cap 150 may further include multiple sealing posts 151-153 extending outward from the cover plate 151. The sealing posts 151-153 are made of soft materials such as silicone or TPE, and their size is slightly larger than the size of the first channel. After each sealing post is inserted into each first channel, the first channel can be sealed, and the injection cap can also be fixedly connected to the first inlet to prevent it from falling off during movement. In some embodiments, the injection cap 150 may further include a limiting edge 155, which is located near the second channel of the container after the injection cap is closed. The limiting edge 155 can engage with the outer surface of the container to further limit the injection cap and ensure that the injection cap seals each first channel.

[0060] In some embodiments, to prevent piston movement during movement or sample dispensing, the microfluidic chip 100 may further include a removable piston limiting block 160. Before sample dispensing, the piston limiting block 160 engages with the container 120 and restricts the position of the piston, preventing the piston from sliding into the container. After sample dispensing is complete, the piston limiting block 160 can be removed before pushing the piston.

[0061] Figure 7 This is a schematic diagram of a piston limiting block structure according to one embodiment of this application. (In conjunction with...) Figure 7 A piston limiting block 160 is inserted between the receiving body 120 and the piston 130 to prevent the piston head from falling. In some embodiments, the piston limiting block includes an operating part 161, a connecting part 162, and a supporting part 163. The operating part 161 provides a grip for the operator to easily remove the piston limiting block; the connecting part 162 extends outward from the operating part 161 and can be inserted into the receiving body; the supporting part 163 extends outward from the operating part and is located at the upper end of the receiving body to support the piston; the supporting part 163 and the connecting part 162 extend outward in parallel in the same direction as the operating part. In some embodiments, the connecting part 162 extends a greater distance than the supporting part 163 to ensure a stable connection between the piston limiting block and the receiving body. In some embodiments, multiple second channels connected to the receiving cavity can be spaced apart, and the connecting part 162 can be inserted between adjacent second channels.

[0062] Figures 8A-8H This is a schematic diagram illustrating different states of a microfluidic chip according to an embodiment of this application. (Reference) Figures 8A-8DTaking the accommodating cavity 121 as an example, before the microfluidic chip is sampled, soluble microspheres 103 are placed in the accommodating cavity 121, the piston plunger 135 is placed in the second channel 1214, the sample inlet cap 150 is placed at the first inlet 1211 of the accommodating cavity, the chip body 110 is disposed on the chip interface 124 of the accommodating body 120 with an interference fit, and the deformable material 140 is located between the chip body 110 and the accommodating body 120. At this time, the deformable material is in an undeformed state.

[0063] refer to Figure 8E-8H After sample addition, the soluble microspheres 103 dissolve upon contact with the sample and mix with it to form a PCR reaction system. The piston 131 pushes the dissolved sample into the reaction zone 111 of the chip body 110. As the dissolved sample enters the reaction zone 111, the gas in the reaction zone 111 can be discharged through the narrow channel 117 via the chip outlet 115 and the exhaust channel 125 of the container. After the piston 131 has pushed all the dissolved sample into the reaction zone, the piston head 136 closes the first channel 1215. The snap-fit ​​portion 138 on the piston 131 snaps into the snap-fit ​​portion 129 of the container. In some embodiments, the deformable material 140 gradually deforms as the piston is pushed. In some embodiments, when the piston 131 pushes the entire sample after the microspheres have dissolved into the chip body 110, the deformable material 140 has not yet sealed the chip inlet of the chip body 110. Further pushing of the piston 131 causes compression between the container and the chip body, and the deformable material 140 continues to deform until the chip inlet is completely sealed. In some embodiments, the thickness of the deformable material 140 changes from 0.5-1.0 mm to 0.2-0.3 mm when it deforms to seal the chip inlet.

[0064] To facilitate the movement of the piston, this application also proposes a sealing device for use in conjunction with it. Figure 9 This is a schematic diagram of a sealing device according to an embodiment of this application. As shown, the sealing device 900 includes a first clamp 910 and a second clamp 920. The first clamp 910 can be used to hold a microfluidic chip, and the second clamp 920 can be used to control the piston movement of the microfluidic chip. In this embodiment, during operation, the sample inlet cap of the microfluidic chip is first removed, and the sample is added to the first inlet. After the sample is added, the microfluidic chip is placed on the first clamp 910; then, the piston limiting block of the microfluidic chip is removed, and the first clamp 910 holding the microfluidic chip is placed on the second clamp 920. The second clamp 920 pushes the piston of the microfluidic chip to push the sample after the microspheres in the accommodating cavity are dissolved into the reaction zone of the microfluidic chip.

[0065] In some embodiments, the first clamp 910 includes a first base 911, a first body 912, and a second body 913. The first body 912 and the second body 913 are located on the first base 911; and a slit 914 is included between the first body 912 and the second body 913, which can be used to accommodate the chip body of the microfluidic chip and can protect the chip body of the microfluidic chip.

[0066] In some embodiments, when the piston is pushed, the first body 912 and the second body 913 can also support the microfluidic chip's housing. As the piston moves, the deformable material deforms to seal the chip inlet. The housing moves onto the first and second bodies, preventing excessive force from damaging the microfluidic chip. In some embodiments, the height of the first seat between the first and second bodies is a first height, and the height between the edges of the housing supported on the first and second bodies is a second height. The height difference between the second height and the first height is equal to or greater than the deformation of the deformable material.

[0067] In some embodiments, the second clamp 920 includes: a second base 921, a first column 922, a second column 923, a support 924, and a pressure block 925. The first column 922 and the second column 923 are disposed on the second base, and the space between the first and second columns can accommodate a first clamp 910 for holding the microfluidic chip. One end of the support 924 is disposed on the second base, and the other end is connected to the first and second columns respectively via a connecting rod 926. The pressure block is disposed between the support and the first / second column and can move relative to the support along the connecting rod 926, thereby pushing the piston of the microfluidic chip. In some embodiments, the height of the first and second columns can be the height at which the microfluidic chip piston pushes all the dissolved microsphere sample into the reaction zone, avoiding excessive pressure from the pressure block that could damage the microfluidic chip.

[0068] In some embodiments, the second clamp 920 may further include a handle 927, which is connected to the pressure block via a bracket. The handle 927 is threadedly connected to the bracket, and rotating the handle controls the movement of the pressure block, which helps maintain pressure uniformity and operational control. In some embodiments, the handle 927 may be a torque handle. A fixed torque value is set to push the piston; that is, rotating the handle controls the pressure block to push the handle. When the torque of the handle reaches the set value, continuing to rotate the handle will not cause the pressure block to follow the handle's rotation, thus preventing the pressure block from excessively pushing the piston and causing chip damage. In some embodiments, the torque value may be 0.7-1.1 Nm.

[0069] Figure 10A and Figure 10BThis is a schematic diagram of a microfluidic chip according to another embodiment of this application. Figure 11 for Figure 10A and Figure 10B An exploded view of the microfluidic chip in the embodiment described. As shown, the microfluidic chip 1000 includes a chip body 1010, a housing 1020, one or more pistons 1030, and a deformable material 1040. Figure 10A and Figure 10B The embodiments are with Figures 1A-1C The parts that are the same or similar to those in the embodiments will not be described again.

[0070] The differences between the two embodiments are explained in detail below. First, in this embodiment, the direction of piston movement is perpendicular to the direction of the chip body. Correspondingly, the chip inlet 1011 and chip outlet 1012 of the reaction region of the chip body 1010 are disposed on the upper surface of the chip body, rather than on the side surface of the chip body. Apart from the difference in the number of reaction regions, the other structures of the chip body in this embodiment are similar to those in the embodiment of FIG1, and will not be described again here.

[0071] In this embodiment, the microfluidic chip includes a chip connector 1050 for bonding the chip body 1010 to the accommodating body 1020. Specifically, the connector 1050 includes an upper retainer 1051 and a lower retainer 1052. The upper retainer 1051 and the lower retainer 1052 interlock and clamp the chip body 1010, thereby tightly bonding the chip body 1010 to the accommodating body 1020. In some embodiments, the lower retainer 1052 is in direct contact with the chip 1010, while a deformable material 1040 is disposed between the upper retainer 1051 and the chip body 1010. Through compression between the upper retainer 1051 and the lower retainer 1052, the deformable material 1040 can undergo partial deformation, sealing the gap between the accommodating body 1020 and the chip body 1010, preventing sample leakage between them. It should be noted that the deformable material 1040 includes two sets of openings, one set for communication between the accommodating cavity and the chip inlet, and the other set for venting from the chip outlet. Although the compression between the upper retainer 1051 and the lower retainer 1052 causes partial deformation of the deformable material 1040, this partial deformation is insufficient to close the two sets of openings, which remain open.

[0072] In some embodiments, the accommodating body 1020 includes a connecting plate 1021 connected to the upper retainer 1051. The connecting plate 1021 includes a retaining ring 1002 extending toward the chip body for engaging with a snap fastener 1001 on the lower retainer 1052, thereby attaching the chip body 1010 to the accommodating body 1020. Other structures of the accommodating body 1020 are similar to those in the embodiment of FIG. 1, and will not be described again here. In some embodiments, the chip body 1010 may also be connected to the accommodating body 1020 in other ways, such as by interference fit or other methods.

[0073] In some embodiments, the structure of piston 1030 is similar to that of the embodiment in FIG. 1, and therefore will not be described again here. In some embodiments, the microfluidic chip includes a set of venting and sealing pistons 1060, which can be connected to the push plate of piston 1030, and the sealing of the chip outlet of chip body 1010 is achieved by the pushing of piston 1030. In some embodiments, venting and sealing piston 1060 includes a slide plate 1061 and a plurality of plug rods 1062, the plurality of plug rods 1062 extending from slide plate 1061 toward chip body 1010. Slide plate 1061 contacts piston 1030, and the movement of piston 1030 causes slide plate 1061 and plurality of plug rods 1062 to slide along accommodating body 1020 toward chip body 1010. In some embodiments, accommodating body 1020 is provided with a groove 1022 for accommodating slide plate, which can limit the sliding process of slide plate 1061. After the piston 1030 pushes the sample after all the microspheres in the accommodating cavity are dissolved into the reaction area of ​​the chip body 1010, the piston continues to push the slide plate 1061 to slide, so that the stop rod 1062 abuts against a set of openings on the deformable material 1040 that correspond to the chip outlet of the chip body 1010, thereby sealing the chip outlet.

[0074] Figure 12 This is a schematic diagram of a deformable material structure according to another embodiment of this application. (In conjunction with...) Figure 12 The deformable material 1040 includes multiple liquid inlet holes 1041 and multiple vent holes 1042, which correspond to the positions of the chip inlet and chip outlet, respectively, so that the chip inlet is connected to the outlet of the accommodating cavity. After the piston 1030 pushes all the dissolved microspheres in the accommodating cavity into the reaction zone, the piston 1030 squeezes the deformable material 1040 to seal the chip inlet; the stopper rod 1062 squeezes the deformable material 1040 to seal the chip outlet.

[0075] In some embodiments, the vent of the deformable material 1040 includes a protruding lip 1043, and the lip 1043 may include a notch 1044. When the piston 1030 pushes the plug rod 1062 to move and contact the lip 1043, the notch can still be used to discharge gas in the reaction zone; as the piston 1030 continues to push, the plug rod 1062 enters the lip, thereby disabling the notch 1043 and sealing the chip outlet.

[0076] Figures 13A-13F This is a schematic diagram illustrating different states of a microfluidic chip according to another embodiment of this application. (See reference...) Figures 13A-13C The microfluidic chip is in its first state (before sample addition). Soluble microspheres 1003 are placed in the containment cavity, the piston plunger is positioned in the second channel, and the sample inlet cap is located at the first inlet of the containment cavity. The chip body 1010 is attached to the containment body 1020 via a connector 1050. Deformable material 1040 is located between the chip body 1010 and the connector 1050. The deformable material is in a partially deformed state. The plunger of the exhaust sealing piston is not fully inserted into the exhaust port of the deformable material, allowing gas to escape normally through the notch on the upper lip of the exhaust port.

[0077] refer to Figure 13D-13F In the second state (i.e., after sample addition), the soluble microspheres 1003 dissolve upon contact with the sample and mix with it to form a PCR reaction system. The piston 1030 pushes the dissolved sample into the reaction zone of the chip body 1010. As the dissolved sample enters the reaction zone, gas in the reaction zone can exit through a narrow channel from the chip outlet. After the piston has pushed all the dissolved sample into the reaction zone, the piston 1030 seals the first channel and the outlet of the accommodating cavity. Furthermore, the piston 1030 cooperates with the deformable material 1040 to seal the chip inlet of the chip body 1010, and the retaining ring on the piston 1030 engages with the retaining protrusion of the accommodating body 1020 to position the piston 1030. In some embodiments, as the piston is pushed, the venting and sealing piston gradually enters the vent hole of the deformable material. After the piston 1030 pushes all the sample after the microspheres have dissolved into the chip body 1010, the plug rod of the exhaust sealing piston is completely inserted into the exhaust hole of the deformable material 1040, causing the exhaust hole to fail and sealing the chip outlet.

[0078] Figure 14A and Figure 14B This is a schematic diagram of a microfluidic chip according to another embodiment of this application. Figure 15This is an exploded view of a microfluidic chip according to another embodiment of this application. As shown, the microfluidic chip includes a chip body 1410, a container 1420, one or more pistons 1430, and a deformable material 1440. The container 1420 is used to accommodate microspheres and receive samples. The pistons 1430 can move into the container 1420 to push the dissolved microsphere sample into the chip body 1410; the deformable material 1440 is disposed between the chip body 1410 and the container 1420 to seal the gap between the chip body 1410 and the container 1420. In this embodiment, the direction of piston movement is parallel to the direction of the chip body. Figure 14A and Figure 14B The embodiments are with Figures 1A-1C The parts that are the same or similar to those in the embodiments will not be described again.

[0079] The differences between the two embodiments are described in detail below. First, the chip body 1410 is connected to the accommodating body 1420 via an attachment 1450. The attachment 1450 clamps the chip body 1410 and is connected to the accommodating body 1420 by an interference fit, thereby clamping the chip body 1410 to the accommodating body 1420.

[0080] In some embodiments, the microfluidic chip 1400 includes a slider 1460 that can contact the push plate of the piston 1430. The slider 1460 includes one or more sealing plugs 1461 disposed on the surface of the slider 1460 facing the container 1420. The slider 1460 contacts the piston 1430; by moving the piston 1430, the slider 1460 can slide along the container toward the chip body, and the sealing plugs 1461 slide accordingly, sealing the exhaust channel of the container 1420. In some embodiments, the container is provided with a groove 1421 for accommodating the slider, located on the surface of the exhaust channel outlet of the container, which can limit the slider's sliding process. After the piston pushes all the dissolved microspheres in the container cavity into the reaction zone, the piston pushes the sliding plate to slide, and the sealing plugs can seal the outlet of the exhaust channel.

[0081] Figures 16A-16D This is a schematic diagram illustrating different states of a microfluidic chip according to another embodiment of this application. (See reference...) Figures 16A-16B The microfluidic chip is in its first state (i.e., before sample addition). Soluble microspheres 1403 are disposed in the accommodating cavity, the piston plunger is disposed in the second channel, and the sample inlet cap is disposed at the first inlet of the accommodating cavity. The chip body 1410 is connected to the accommodating body via an attachment 1450, and a deformable material 1440 is located between the accommodating body 1420 and the attachment 1450. In some embodiments, the deformable material 1440 is not in a deformable state. In some embodiments, the slider is in contact with the piston, and the sealing plug 1461 on the slider is away from the exhaust channel outlet on the accommodating body, allowing gas to be discharged normally through the exhaust channel.

[0082] refer to Figure 16C-16D In the second state (i.e., after sample addition), the soluble microspheres 1403 are dissolved upon contact with the sample and mixed with it to form the PCR reaction system. The piston 1430 pushes the dissolved sample into the reaction zone of the chip body 1410. As the dissolved sample enters the reaction zone, gas from the reaction zone passes through a narrow channel from the chip outlet into the exhaust channel of the container 1420 and exits through the exhaust channel outlet. With the piston 1430 being pushed, the deformable material 1440 gradually deforms. In some embodiments, when the piston 1430 has completely pushed the dissolved sample into the chip body 1410, the deformable material 1440 has not yet sealed the chip inlet of the chip body 1410; further pushing of the piston 1430 causes the container 1420 and the attachment 1450 to continue to compress, causing the deformable material 1440 to continue to deform and seal the chip inlet. In this embodiment, as the piston 1430 is pushed, the sealing plug of the slider 1460 gradually approaches the outlet of the exhaust channel of the container 1420. After the piston 1430 pushes all the sample after the microspheres are dissolved into the chip body 1410, the sealing plug on the slider 1460 completely covers the outlet of the exhaust channel on the container 1420, and the exhaust channel cannot discharge gas, thus achieving a seal on the chip outlet.

[0083] The microfluidic chip of this application can seal the chip inlet and outlet of the reaction zone after the sample dissolved by the microspheres enters the reaction zone through different sealing methods, forming a fully enclosed system. This can prevent external impurities from affecting the PCR reaction, improve the specificity and accuracy of detection, and also make the evaporation rate of the liquid in the PCR reaction <5% at high temperature (such as 95℃).

[0084] This application also proposes a method for operating microfluidic chips. Figure 17 The following is a schematic diagram of the operation process of a microfluidic chip according to an embodiment of the present application, which includes the following steps: For the various embodiments of the present invention as described herein, in step 1710, a sample after nucleic acid extraction is added from the first inlet of the microfluidic chip's container. The sample after nucleic acid extraction can dissolve the soluble microspheres disposed in the container cavity, thereby forming a reaction system suitable for PCR.

[0085] In step 1720, the piston of the microfluidic chip is pushed to introduce the dissolved microsphere sample into the reaction zone of the microfluidic chip. In some embodiments, after the piston has completely pushed the reaction solution into the reaction zone, the piston is further pushed, and a deformable material can seal the chip inlet of the reaction zone.

[0086] In step 1730, the microfluidic chip is transferred into a PCR instrument, which performs PCR reaction and optical detection on the sample. In some embodiments, the detection time of the PCR instrument is within 2 hours, within 1.5 hours, or within 1 hour.

[0087] The above embodiments are for illustrative purposes only and are not intended to limit the invention. Those skilled in the art can make various changes and modifications without departing from the scope of the invention. Therefore, all equivalent technical solutions should also fall within the scope of the invention.

Claims

1. A microfluidic chip, comprising: A chip body includes a flat reaction region, the reaction region including a chip inlet and a chip outlet, and an elongated channel between the reaction region and the chip outlet; A container includes a cavity for accommodating soluble microspheres, the cavity including a first inlet, a second inlet, and a first outlet, the first inlet being for receiving a sample, and the first outlet being connected to the chip inlet of the reaction zone; A piston configured to push a sample of dissolved microspheres from the accommodating cavity into the reaction zone through the second inlet; as well as A deformable material that, after deformation, seals the chip inlet of the reaction zone.

2. The microfluidic chip according to claim 1, wherein the depth of the reaction zone is 200-450 μm and the volume is 10-15 μl, and the thermal conductivity of at least a portion of the material of the chip body is such that the average heating rate of the reaction zone is ≥15 °C / second when the heater temperature is 95 °C.

3. The microfluidic chip according to claim 1, wherein the cross-sectional area of ​​the elongated channel is 0.07-0.2 mm². 2 The length is 15-25mm.

4. The microfluidic chip according to claim 1, wherein the accommodator includes a first channel connected to the first inlet; a second channel connected to the second inlet, the second channel being adapted to the piston, the first channel being connected to the second channel, and the piston closing the first channel after passing through the first channel.

5. The microfluidic chip according to claim 1, wherein the accommodator includes a removable injection cap that is adapted to and closes the first inlet.

6. The microfluidic chip according to claim 1, wherein when the piston pushes the dissolved sample of the microspheres in the accommodating cavity into the reaction zone, the deformable material gradually deforms.

7. The microfluidic chip according to claim 1, wherein when the piston pushes the sample after all the dissolved microspheres in the accommodating cavity into the reaction zone, the deformable material has not yet sealed the chip inlet of the chip body.

8. The microfluidic chip according to claim 7, wherein when the piston pushes the sample after all the dissolved microspheres in the accommodating cavity into the reaction zone and continues to move, the deformable material continues to deform and seal the chip inlet.

9. The microfluidic chip according to claim 1, wherein the accommodator includes an exhaust channel connected to the chip outlet, and the exhaust channel is provided with a water-repellent seal.

10. The microfluidic chip according to claim 1, wherein, After the piston pushes the sample containing all the dissolved microspheres in the accommodating cavity into the reaction zone, the deformable material seals the chip outlet after deformation.

11. The microfluidic chip according to claim 1, further comprising a sealing plug, wherein, After the piston pushes the sample containing all the dissolved microspheres in the accommodating cavity into the reaction zone, the sealing plug moves and seals the chip outlet.

12. The microfluidic chip according to claim 1, wherein the direction of movement of the piston is parallel to the direction of the chip body.

13. The microfluidic chip according to claim 12, wherein the accommodator includes a chip interface for accommodating the chip body, the chip body and the chip interface are interference-fitted, and the deformable material is disposed within the chip interface.

14. The microfluidic chip according to claim 1, wherein the direction of movement of the piston is perpendicular to the direction of the chip body.

15. The microfluidic chip of claim 14, wherein the chip body is attached to the accommodating body, the chip inlet of the chip body corresponds to the first outlet of the accommodating cavity, and the deformable material is disposed between the chip inlet and the first outlet.

16. The microfluidic chip of claim 14, further comprising an exhaust sealing piston, wherein the deformable material includes an exhaust port communicating with the chip outlet, wherein when the piston pushes the sample after all the dissolved microspheres in the accommodating cavity into the reaction zone and the deformable material seals the chip inlet after deformation, the exhaust sealing piston seals the exhaust port of the deformable material.

17. The microfluidic chip of claim 16, wherein the vent of the deformable material includes a protruding lip with a notch for venting, and the vent sealing piston enters the lip to disable the notch on the lip.

18. The microfluidic chip of claim 1, further comprising one or more additional pistons, the chip body including one or more additional flat reaction zones, the accommodating body including one or more additional accommodating cavities, the one or more additional pistons being configured to push a sample of dissolved microspheres from the corresponding one or more additional accommodating cavities into the corresponding one or more additional reaction zones.

19. The microfluidic chip according to claim 1, wherein the sample is a sample processed by nucleic acid extraction, and the sample is dissolved in the microspheres to obtain a system suitable for PCR reaction.

20. The microfluidic chip according to claim 1, wherein the PCR reaction system after the microspheres are dissolved is used for HPV detection.

21. A sealing device for use with a microfluidic chip according to any one of claims 1-20, comprising: The first clamp includes: The first body; and The first body and the second body are located on the first seat; Wherein, a slit is included between the first body and the second body, the slit being adapted to accommodate the chip body of the microfluidic chip, and the first body and the second body being adapted to receive the accommodating body of the microfluidic chip; The second clamp includes: The second body; The first and second pillars located on the second base; The bracket located on the second body; and Pressure block connected to the bracket; The space between the first column and the second column is adapted to accommodate the first clamp; the pressure block is configured to move relative to the support, thereby actuating the piston of the microfluidic chip.

22. The sealing device according to claim 21, wherein the height between the first seat and the first body and the second body is a first height, the height between the chip body of the microfluidic chip and the edge of the accommodating body that is supported on the first body and the second body is a second height, the first height is greater than the second height, and the height difference between the first height and the second height is equivalent to or greater than the deformation of the deformable material.

23. A method of operating a microfluidic chip according to any one of claims 1-20, comprising: A sample after nucleic acid extraction is added from the first inlet of the container of the microfluidic chip, and the sample after nucleic acid extraction dissolves the microspheres in the container cavity to form a reaction system suitable for PCR. The piston of the microfluidic chip pushes the sample after dissolving the microspheres into the reaction zone of the microfluidic chip; as well as The microfluidic chip was then transferred into the PCR instrument.

24. The method of claim 23, further comprising: After pushing the piston of the microfluidic chip to push the sample after dissolving the microspheres into the reaction zone of the microfluidic chip, the piston is further pushed to seal the chip inlet of the reaction zone.

25. The method according to claim 23, wherein the detection time of the PCR instrument is within 2 hours, within 1.5 hours, or within 1 hour.

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