Microfluidic chip and method of using the same, microfluidic system and method of manufacturing conductive cover plate

Abstract

A microfluidics chip (304) and its method of use, a microfluidics system, and a method of manufacturing a conductive cover plate (4) are disclosed. The chip (304) includes an extraction assembly (1) and an amplification assembly (2). The extraction assembly (1) includes a lysis chamber (100), a first valve chamber (101), a washing chamber (102, 103), and a second valve chamber (104), which are connected in series. The amplification assembly (2) includes a chip substrate (6), a barrier layer (5), and a conductive cover plate (4). The barrier layer (5) is located between the chip substrate (6) and the conductive cover plate (4). The barrier layer (5), the chip substrate (6), and the conductive cover plate (4) define an elution chamber (21), a liquid path chamber (22), and an amplification chamber (23), which are connected in series. The elution chamber (21) and the second valve chamber (104) are connected via a first channel (24). The plate (6) is provided with an electrode array (7), the conductive cover plate (4) is provided with a cavity (400) with an open bottom on the side closer to the chip substrate (6), the elution chamber (21) includes the cavity (400), the electrode array (7) is located directly below the elution chamber (21), the liquid path chamber (22), and the amplification chamber (23), the amplification assembly (2) further includes a storage chamber (20) for storing a surfactant, the storage chamber (20) and the elution chamber (21) are in communication with each other via a second channel (25), the liquid path chamber (22) includes a liquid storage cavity (220), and the liquid storage cavity (220) is adjacent to the elution chamber (21), and the thickness of the cavity (400) in the thickness direction of the microfluidics chip (304) is 0.6 mm to 2 mm.

Description

[Technical Field]

[0001] The present invention relates to the technical field of microfluidics, in particular to a microfluidics chip and its use method, a microfluidics system, and a manufacturing method for a conductive cover plate. This application is based on the Chinese invention patent application filed on October 12, 2022 with application number CN202211247595.1, filed on July 11, 2023 with application number CN202310848619.7, and the Chinese utility model patent application filed on October 12, 2022 with application number CN202222690774.4, the contents of which are incorporated herein by reference. [Background technology]

[0002] Digital microfluidics (DMF) is a branch of microfluidics technology that can control single droplets of microliter to nanoliter size. Based on the electrowetting-on-dielectric (EWOD) phenomenon, digital microfluidics technology can manipulate single droplets on electrode arrays through electrowetting forces. Its electrically actuated capability and small footprint make it a promising technology for point-of-care diagnostics.

[0003] There are two types of PCR methods in digital microfluidics systems: stationary PCR, which is based on the time domain, and shuttling PCR, which is based on the space domain. In stationary PCR experimental setups, PCR reaction droplets remain on electrodes for in situ PCR reactions, and thermal cycling is achieved by controlling the heating time. The heater can be a large external heater that heats and cools the entire chip, or it can be an on-chip heater with a small heating volume. In real-time processes using PCR for qualitative and quantitative analysis in microfluidics, first, the reagents need to be heated to 95°C. Second, most PCR methods use a fluorescent method, which requires the cooperation of an optical module. At this temperature, the reagents are prone to evaporating and generating bubbles, which affects the morphology of the reagents and makes it impossible to read the amplification curve.

[0004] Existing microfluidic chips include a conductive cover plate and a chip substrate, with a Teflon hydrophobic layer on the side of the conductive cover plate closest to the chip substrate and an expanded PTFE film on the side of the chip substrate closest to the conductive cover plate, which reduces or prevents the generation of air bubbles when the microfluidic chip performs an amplification reaction. However, this method is costly, relatively complicated to produce and manufacture, and not suitable for mass production.

[0005] In addition, there are microfluidic chips in which a hydrophilic layer is applied to the side of the conductive cover plate closest to the chip substrate, and a hydrophobic layer is provided on the side of the chip substrate closest to the conductive cover plate. However, with this solution, once bubbles are generated, the sample reagent is easily pushed by the generated bubbles, which can cause the sample to move and interfere with observation.

[0006] Conventional integrated microfluidic chips include a lysing chamber, a washing chamber, an elution chamber, a liquid channel chamber, and an amplification chamber, all of which are connected in series. The lysing chamber contains a lysing solution, the washing chamber contains a cleaning solution, and the elution chamber contains an elution solution. The elution chamber and the liquid channel chamber are typically separated by a spacer such as a paraffin valve, and the substrate directly below the elution chamber typically does not have an electrode array. However, in this microfluidic chip, the method of storing the elution solution is quite tedious. The time required to sequentially heat and melt the paraffin valves reduces detection efficiency. Furthermore, because the substrate directly below the elution chamber typically does not have an electrode array, it is impossible to transfer all eluted sample droplets to the liquid channel chamber, resulting in inaccurate sample detection.

[0007] To move all the eluted sample droplets into the liquid channel chamber, existing microfluidic chips typically use air pumps or gravity to move the sample droplets into the liquid channel chamber. Both operations are tedious and complicated, and the latter cannot perform more advanced operations such as liquid separation.

[0008] One type of existing microfluidics chip includes a liquid storage electrode, a liquid separation electrode, and a liquid stationary electrode, which are arranged in order, with the liquid storage electrode being wider than the liquid stationary electrode and the liquid separation electrode being narrower than the liquid stationary electrode. Before liquid separation, the sample droplet covers the entire liquid storage electrode and part of the liquid separation electrode. When transferring liquid, first, the liquid separation electrode is energized, and the sample droplet occupies most of the liquid storage electrode, covers the liquid separation electrode, and occupies most of the liquid stationary electrode. At this time, the liquid separation electrode is turned off, and both the liquid storage electrode and the liquid stationary electrode are energized to rapidly split the sample droplet. Finally, the liquid storage electrode and the liquid stationary electrode are turned off, and the droplet separated from the liquid storage electrode is on the liquid stationary electrode. However, when separating sample droplets in this manner, if the power supply to the liquid separation electrode is turned off, the volume of the sample droplets on the liquid separation electrode becomes too large, causing the droplets to split slowly during liquid separation. As a result, the droplets split unevenly, and the amount of droplets on the liquid fixed electrode becomes insufficient to meet the experimental requirements, or the amount of droplets on the liquid fixed electrode becomes too large, affecting the accuracy of the experiment. Summary of the Invention [Problem to be solved by the invention]

[0009] The first object of the present invention is to provide a microfluidics chip that allows for easy storage and transfer of eluent, can improve detection efficiency, and can sufficiently transfer sample droplets in the elution chamber into the liquid channel chamber.

[0010] A second object of the present invention is to provide a microfluidics chip that can split sample droplets uniformly and accurately when separating liquids.

[0011] A third object of the present invention is to provide a microfluidic chip that reduces the influence of air bubbles on detection results in the PCR process.

[0012] A fourth object of the present invention is to provide a method for fabricating a conductive cover plate that is applied to the above-mentioned microfluidic chip.

[0013] A fifth object of the present invention is to provide a method for using the above microfluidics chip.

[0014] A sixth object of the present invention is to provide a microfluidics system including the above-mentioned microfluidics chip. [Means for solving the problem]

[0015] In order to achieve the first object, the microfluidic chip according to the present invention includes an extraction assembly and an amplification assembly. The extraction assembly includes a lysis chamber, a first valve chamber, a washing chamber, and a second valve chamber, which are sequentially connected to each other. The amplification assembly includes a chip substrate, a barrier layer, and a conductive cover plate. The barrier layer is located between the chip substrate and the conductive cover plate. The barrier layer, the chip substrate, and the conductive cover plate define an elution chamber, a liquid path chamber, and an amplification chamber, which are sequentially connected to each other. The elution chamber and the second valve chamber are separated by a first chamber. The microfluidics chip has a microfluidic chip and a microchannel chamber, and the microfluidic chip is connected to the microfluidic chip through a second channel. The microfluidic chip has a chip substrate having an electrode array on the chip substrate. The conductive cover plate has a cavity with an open bottom on the side closer to the chip substrate. The elution chamber includes the cavity. The electrode array is located directly below the elution chamber, the liquid path chamber, and the amplification chamber. The amplification assembly further includes a storage chamber for storing a surfactant. The storage chamber and the elution chamber are connected to each other through a second channel. The liquid path chamber includes a liquid storage cavity. The liquid storage cavity is adjacent to the elution chamber. The thickness of the cavity is 0.6 mm to 2 mm in the thickness direction of the microfluidics chip.

[0016] In a further aspect, the electrode array includes an elution electrode region and a first liquid storage electrode region adjacent to each other, the elution electrode region including at least one elution electrode, the elution electrode being located within the elution chamber when projected onto the chip substrate, the first liquid storage electrode region including one first transition electrode adjacent to each other and a first liquid storage electrode group, the first transition electrode spanning the elution chamber and the liquid storage cavity when projected onto the chip substrate, and the first transition electrode being located between the elution electrode region and the first liquid storage electrode group, the first liquid storage electrode group being located within the liquid storage cavity.

[0017] In a further embodiment, the first liquid storage electrode group includes at least two first liquid storage electrodes, the plurality of first liquid storage electrodes being arranged in order in a direction from the elution chamber to the liquid path chamber, the area of ​​the next first liquid storage electrode being larger than the area of ​​the previous first liquid storage electrode, and the previous first liquid storage electrode being closer to the first transition electrode than the next first liquid storage electrode.

[0018] In a further embodiment, each first liquid storage electrode includes a first side edge and a second side edge extending in a direction from the elution chamber to the liquid path chamber, the first side edge and the second side edge being opposite each other, and the distance between the first side edge and the second side edge of the next first liquid storage electrode being greater than the distance between the first side edge and the second side edge of the previous first liquid storage electrode.

[0019] In a further embodiment, the first side edges of all the first liquid storage electrodes are arranged on the same straight line, and the second side edges of all the first liquid storage electrodes are arranged on the same straight line.

[0020] In a further embodiment, the first transition electrode and the first liquid storage electrode adjacent to the first transition electrode have first arc-shaped portions that fit together, with openings of the first arc-shaped portions facing toward the first transition electrode, and two adjacent first liquid storage electrodes have second arc-shaped portions that fit together, with openings of the second arc-shaped portions facing toward the first transition electrode.

[0021] In a further aspect, at least one first arcuate segment is provided within the first arcuate portion, with an opening in the first arcuate segment facing the first transition electrode, and at least one second arcuate segment is provided within the second arcuate portion, with an opening in the second arcuate segment facing the first transition electrode.

[0022] In a further embodiment, the conductive cover plate includes a plate and a cover, the cavity having an open top, and the cover covering the open top of the cavity.

[0023] In order to achieve the second object, the microfluidics chip according to the present embodiment includes an amplification assembly, the amplification assembly including a chip substrate, a barrier layer, and a conductive cover plate, the chip substrate being disposed opposite the conductive cover plate, the chip substrate including a base plate, an electrode array, and an insulating layer, the electrode array being located on the base plate, the insulating layer covering the electrode array and being close to the conductive cover plate, the barrier layer being located between the chip substrate and the conductive cover plate, and a receiving portion being formed between the barrier layer, the chip substrate, and the conductive cover plate. The electrode array corresponds to the accommodation section, and includes a second liquid storage electrode region, a liquid separation electrode region, and a liquid fixing electrode region adjacent to each other in this order, the width of the liquid fixing electrode region and the width of the second liquid storage electrode region are all greater than the width of the liquid separation electrode region, the liquid fixing electrode region includes at least one liquid fixing electrode, the second liquid storage electrode region includes at least one second liquid storage electrode, and the liquid separation electrode region includes a liquid separation section, and the liquid separation section includes a first electrode and a second electrode arranged adjacent to each other in the width direction of the liquid separation section, and the area of ​​the second electrode is smaller than the area of ​​the liquid fixing electrode.

[0024] In a further embodiment, the liquid separation electrode region further includes a liquid transport portion, the liquid transport portion including at least one liquid transport electrode, the liquid transport portion being located between the liquid separation portion and the second liquid storage electrode region.

[0025] In a further embodiment, the width direction of the liquid fixed electrode intersects with the width direction of the liquid transport section, the width direction of the first end of the liquid separation section is parallel to the width direction of the liquid transport section and the first end of the liquid separation section is close to the liquid transport section, the width direction of the second end of the liquid separation section is parallel to the width direction of the liquid fixed electrode area and the second end of the liquid separation section is close to the liquid fixed electrode area.

[0026] In a further embodiment, the width direction of the liquid fixing electrode region and the width direction of the liquid transporting portion are perpendicular to each other.

[0027] In a further embodiment, the second liquid storage electrode region includes two or more second liquid storage electrodes adjacent to each other in sequence, where the two adjacent second liquid storage electrodes have first arc-shaped edges that fit together, with the openings of the first arc-shaped edges facing toward the liquid separation electrode region, and the liquid fixing electrode region includes two or more liquid fixing electrodes adjacent to each other in sequence, where the two adjacent liquid fixing electrodes have second arc-shaped edges that fit together, with the openings of the second arc-shaped edges facing toward the liquid separation electrode region.

[0028] In a further embodiment, a third arcuate edge is provided within the first arcuate edge, with an opening in the third arcuate edge facing the liquid separation electrode region, and a fourth arcuate edge is provided within the second arcuate edge, with an opening in the fourth arcuate edge facing the liquid separation electrode region.

[0029] In a further aspect, the area of ​​the first electrode is greater than the area of ​​the second electrode.

[0030] In a further embodiment, the shape of the second electrode includes an isosceles triangle, the hypotenuse of the second electrode is adjacent to the first electrode, and the liquid transport portion and the liquid fixing electrode region are each adjacent to two adjacent sides of the second electrode.

[0031] In a further embodiment, the second liquid storage electrode region further includes two second transition electrodes, and the liquid transport electrode adjacent to the second liquid storage electrode is located between the two second transition electrodes, and each second transition electrode is adjacent to the second liquid storage electrode and the liquid transport electrode, respectively.

[0032] In a further embodiment, the liquid separating portion includes a first tooth-shaped portion, a second tooth-shaped portion, and a third tooth-shaped portion, the first tooth-shaped portion being located on an edge of the liquid separating portion close to the liquid fixation electrode region and engaging with an edge of the liquid fixation electrode region, the second tooth-shaped portion being located on an edge of the liquid separating portion close to the liquid transfer portion and engaging with an edge of the liquid transfer portion, and the third tooth-shaped portion being located on an edge of the second electrode close to the first electrode and engaging with the first electrode.

[0033] In order to achieve the third object, a microfluidics chip according to the present invention includes an amplification assembly, the amplification assembly including a chip substrate, a conductive cover plate, a barrier layer, and a first hydrophobic layer, the chip substrate being disposed opposite the conductive cover plate, the chip substrate including a base plate, an electrode array, and an insulating layer, the first hydrophobic layer being located on a side of the insulating layer closer to the conductive cover plate, the electrode array being located on the base plate, the insulating layer covering the electrode array, the barrier layer being located between the chip substrate and the conductive cover plate, A liquid path chamber and an amplification chamber are formed between the chip substrate and the conductive cover plate, and are connected to each other; the liquid path chamber, the amplification chamber, and the electrode array are correspondingly arranged; the conductive cover plate includes a liquid path region and an amplification region connected to each other; the liquid path region corresponds to the liquid path chamber, and the amplification region corresponds to the amplification chamber; the amplification assembly further includes adjacent second hydrophobic and hydrophilic layers; the hydrophilic layer is located on a side of the amplification region closer to the chip substrate; and the second hydrophobic layer is located on a side of the liquid path region closer to the chip substrate; and the second hydrophobic and hydrophilic layers are arranged opposite the first hydrophobic layer.

[0034] In a further aspect, the barrier layer comprises a mixture of adhesive and plastic beads, with the spacing between the chip substrate and the conductive cover plate equal to the diameter of the plastic beads.

[0035] In a further embodiment, the ratio of the density of the adhesive to the density of the plastic beads is 95% or more.

[0036] In a further embodiment, the microfluidics chip includes an extraction assembly, the extraction assembly being in communication with an amplification assembly, the extraction assembly including a lysing chamber, a washing chamber, and an elution chamber, which are in communication with each other in sequence, the lysing chamber, the washing chamber, and the elution chamber being separated by a paraffin valve, the microfluidics chip further including a heating unit provided in the chip substrate, the heating unit including a first heating wire and a second heating wire provided in the base plate, the first heating wire corresponding to the paraffin valve, and the second heating wire corresponding to the amplification chamber.

[0037] In order to achieve the above-mentioned fourth object, the manufacturing method of the conductive cover plate according to the present invention is applicable to any of the above-mentioned microfluidic chips, and includes the steps of applying a first hydrophobic layer on the conductive cover plate, attaching the conductive cover plate with the applied first hydrophobic layer to a fixing jig, and erasing the first hydrophobic layer on the surface of the amplification region with an erasing tool.

[0038] In a further embodiment, the surface of the amplification region from which the first hydrophobic layer has been removed is subjected to a hydrophilization treatment.

[0039] In a further aspect, the fixture exposes only the amplification region of the conductive cover plate.

[0040] To achieve the above-mentioned fifth objective, the present invention provides a method for using the microfluidic chip described in the above-mentioned first objective, which includes the steps of controlling a test sample droplet containing magnetic beads using a magnetic attraction device to move through the dissolution chamber and the washing chamber in sequence into the elution chamber, controlling the magnetic beads using the magnetic attraction device to move back and forth between the storage chamber and the elution chamber, and powering on the electrode array to cause the sample droplet in the elution chamber to enter the liquid storage cavity.

[0041] To achieve the sixth object, the microfluidics system of the present invention includes the microfluidics system of the microfluidics chip described above, and further includes a driving circuit and a control terminal, the control terminal being electrically connected to the driving circuit and used to send control commands to the driving circuit, and the driving circuit being electrically connected to the electrode array and used to control changes in the current-carrying state of the electrode array.

[0042] In a further embodiment, the microfluidics system further includes a magnetic attraction device and a fluorescence detection device, wherein the magnetic attraction device is used to control the sample to move within the extraction assembly, and the fluorescence detection device is used to detect the results of amplifying the sample within the amplification section. [Effects of the Invention]

[0043] The thickness of the cavity wall of the microfluidic chip of the present invention is 0.6 mm to 2 mm. Due to its relatively high surface tension, the eluent without surfactant is blocked by the cavity sidewall and cannot enter the liquid storage cavity or storage chamber. The liquid storage cavity and elution chamber do not need to be separated by a spacer such as a paraffin valve, which simplifies operation and manufacturing. Furthermore, an electrode array is installed below the elution chamber, allowing all of the liquid in the elution chamber to enter the liquid storage cavity through electrowetting, simplifying operation and enabling a variety of advanced operations.

[0044] In the present invention, the provision of the first transition electrode allows the sample droplets to be transferred more conveniently and smoothly from the elution chamber to the liquid storage cavity, and prevents the sample droplets from being blocked by the peripheral wall of the elution chamber and from being unable to transfer completely to the liquid storage cavity due to the dual effects of the gap between the electrodes.

[0045] In the present invention, the area of ​​the next first liquid storage electrode is larger than the area of ​​the previous first liquid storage electrode, so that the liquid storage cavity can temporarily store the test sample droplets sufficiently and meet the needs of detecting large amounts of samples.

[0046] In the present invention, the distance between the first side edge and the second side edge of the next first liquid storage electrode is larger than that of the previous first liquid storage electrode, which can effectively reduce the volume of the chip and make the structure more compact.

[0047] In the present invention, the first side edges of all the first liquid storage electrodes are arranged on the same straight line, and the second side edges of all the first liquid storage electrodes are arranged on the same straight line, thereby increasing the movement speed of the sample droplets.

[0048] In the present invention, the first arc-shaped portion, the first arc-shaped segment, the second arc-shaped portion, and the second arc-shaped segment are provided, thereby enabling the sample droplet to move more smoothly between two adjacent electrodes.

[0049] The present invention provides a liquid separation section with a first electrode and a second electrode arranged side by side across the width of the liquid separation section. When one of the electrodes is energized, the volume of the sample droplet in the liquid separation electrode region is effectively reduced, preventing the electrode droplet from suddenly splitting and breaking apart. When the electrode in the liquid separation region is not energized, the sample droplet splits quickly, resulting in a more accurate droplet size, which meets the experimental requirements.

[0050] When the liquid separation unit of the present invention is located directly adjacent to the second liquid storage electrode, the number of droplets separated by the second liquid storage electrode increases, which affects the volume of sample droplets separated on the liquid fixed electrode.The liquid transfer unit provided in this embodiment can increase the length of the liquid separation electrode area, thereby making droplet splitting more uniform and accurate.

[0051] The width direction of the liquid fixing electrode and the width direction of the liquid transport electrode of the present invention are perpendicular to each other, which makes the arrangement of the electrode array more rational and effectively reduces the size of the microfluidic chip.

[0052] In the present invention, the area of ​​each liquid retention electrode is designed and the required area of ​​the sample droplet is calculated as required, thereby controlling how many liquid retention electrodes the sample droplet covers and obtaining a more accurate size of sample droplet. Furthermore, two adjacent liquid retention electrodes have second arc-shaped edges with openings facing the liquid separation electrode region, thereby accelerating the movement speed of the sample droplet on the liquid retention electrode from the liquid separation electrode region toward the liquid retention electrode region. The number of second liquid storage electrodes is two or more, and the two adjacent second liquid storage electrodes are interdigitated with each other and have first arc-shaped edges with openings facing the liquid separation electrode region, thereby accelerating the movement speed of the sample droplet located on the second liquid storage electrode region from the liquid separation electrode region toward the second liquid storage electrode region. In this way, when separating the liquid, multiple second liquid storage electrodes are energized to promote the splitting of the sample droplet.

[0053] In the present invention, the first arcuate edge is provided with a third arcuate edge that opens opposite the liquid separation electrode region, thereby allowing the sample droplet located in the second liquid storage electrode region to move more smoothly, and the second arcuate edge is provided with a fourth arcuate edge that opens opposite the liquid separation electrode region, thereby allowing the sample droplet located in the liquid fixing electrode region to move more smoothly.

[0054] In the present invention, the area of ​​the first electrode is larger than the area of ​​the second electrode, so when the first electrode is turned off and the second electrode remains energized, there will be fewer sample droplets on the second electrode, and when the second electrode is turned off, the sample droplets on the second electrode will immediately break up.

[0055] The second electrode of the present invention has an isosceles triangular shape, which causes the sample droplets to break up more uniformly.

[0056] In the present invention, by providing transition electrodes on both sides of the liquid transfer electrode adjacent to the second liquid storage electrode, it is possible to prevent the sample droplet from moving from the second liquid storage electrode to the liquid transfer electrode and covering the outside of the liquid transfer electrode, thereby enabling complete control of the movement of the sample droplet.

[0057] In the present invention, the provision of the first toothed portion, the second toothed portion, and the third toothed portion allows the sample droplet to move more smoothly as it passes through the liquid transport electrode, the liquid separation electrode, and the liquid fixing electrode in sequence.

[0058] Aqueous reagents and test samples have strong wettability to hydrophilic surfaces, resulting in strong adsorption of the reagents to the hydrophilic surface. The conductive cover plate with a hydrophilic layer of the present invention has excellent air-repelling properties, allowing the aqueous reagents and test samples to diffuse more effectively within the amplification region. The chip substrate is coated with a first hydrophobic coating, which reduces or even prevents the generation of bubbles during the amplification reaction. After heating the chip substrate, bubbles generated by the aqueous reagents and test samples are quickly removed, preventing the reagents from continuously evaporating and affecting observation. The second hydrophobic layer in the liquid channel region surrounds the hydrophilic layer, allowing the reagents to stably perform the amplification reaction in the amplification region and preventing bubbles from displacing the reagents and affecting observation.

[0059] In the present invention, the adhesive is mixed with plastic beads of a predetermined diameter, allowing the adhesive to achieve a sealing and bonding effect while accurately limiting the height between two flat surfaces. Compared to conventional methods that use gaskets or adhesives to limit the height, this method reduces the difficulty and time required for processing.

[0060] Since the density of the plastic beads of the present invention is close to that of the adhesive, the plastic beads can be dispersed uniformly in the adhesive, and uneven settling of the particles can be avoided.

[0061] The present invention effectively simplifies the operation procedure by combining the extraction assembly and the amplification assembly. The paraffin valve can effectively separate the pre-stored reagents between different chambers in the extraction assembly and prevent contamination between different reagents. The heating wire can be placed in the base plate, which can effectively reduce the volume of the microfluidics chip.

[0062] The present invention can effectively improve the hydrophilicity of the amplification region by subjecting the surface of the amplification region from which the hydrophobic coating has been removed to a hydrophilic treatment.

[0063] In the present invention, the reciprocating motion of the magnetic beads mixes the surfactant in the storage chamber with the sample droplets thoroughly and uniformly, reducing the surface tension of the sample droplets and facilitating their movement.

[0064] As described above, the microfluidic chip of the present invention can stabilize sample reagents in the amplification region and facilitate observation while reducing the risk of bubbles being generated during the PCR process. This chip has a simple and compact structure, convenient eluent storage, a larger capacity, improved detection efficiency, and ensures that sample droplets in the elution chamber are fully transported to the liquid channel chamber. Furthermore, during the liquid separation process, sample droplets can be controlled to split uniformly, and the size of the split sample droplets can be more precisely controlled. [Brief explanation of the drawings]

[0065] [Figure 1] FIG. 1 is a structural diagram of an embodiment of the microfluidic chip of the present invention at a first viewing angle. [Figure 2] FIG. 1 is a structural diagram of an embodiment of the microfluidic chip of the present invention at a second viewing angle, in which the conductive cover plate is divided into a liquid channel region and an amplification region. [Figure 3] FIG. 1 is a structural diagram of the division of each cavity region at a second viewing angle in an embodiment of the microfluidic chip of the present invention. [Figure 4] 1 is a cross-sectional view of an amplification assembly of an embodiment of a microfluidic chip of the present invention. [Figure 5] FIG. 1 is a cross-sectional view of an extraction assembly of an embodiment of a microfluidic chip of the present invention. [Figure 6] FIG. 1 shows a test sample placed between two hydrophobic layers in the amplification region of a conventional microfluidic chip. [Figure 7] FIG. 1 shows a test sample in the amplification region of an embodiment of a microfluidic chip of the present invention. [Figure 8] FIG. 2 is a partial cross-sectional view of an embodiment of the microfluidic chip of the present invention, including an elution chamber, a first channel, and a liquid path chamber. [Figure 9] FIG. 1 is a partial cross-sectional view of an embodiment of the microfluidic chip of the present invention including an elution chamber and a storage chamber. [Figure 10] FIG. 2 is a schematic plan view of an elution electrode region and a first liquid storage electrode region in an embodiment of the microfluidic chip of the present invention. [Figure 11] FIG. 2 is a plan view of an electrode array according to an embodiment of the present invention. [Figure 12] FIG. 12 is an enlarged view of part A in FIG. [Figure 13] FIG. 2 is a first state diagram of a sample droplet on an electrode array according to an embodiment of the present invention. [Figure 14] FIG. 10 is a second state diagram of a sample droplet on an electrode array according to an embodiment of the present invention. [Figure 15] FIG. 10 is a third state diagram of a sample droplet on an electrode array according to an embodiment of the present invention. [Figure 16] FIG. 10 is a fourth state diagram of a sample droplet on an electrode array according to an embodiment of the present invention. [Figure 17] FIG. 1 is a structural block diagram of an embodiment of a microfluidics system of the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0066] As shown in Figures 1 to 7, the microfluidics chip 304 of this embodiment includes an extraction cover plate 3, a conductive cover plate 4, a barrier layer 5, and a chip substrate 6, where the extraction cover plate 3 is adjacent to the conductive cover plate 4, and the conductive cover plate 4 and the extraction cover plate 3 are both arranged opposite the chip substrate 6, and the extraction cover plate 3 and the chip substrate 6 are combined to form an extraction assembly 1, which includes a dissolving chamber 100, a first valve chamber 101, a first cleaning chamber 102, a second cleaning chamber 103, and a second valve chamber 104 that are connected to each other in sequence, and the first valve chamber 101 and the second valve chamber 104 are filled with paraffin. The barrier layer 5, the chip substrate 6, and the conductive cover plate 4 are combined to form the amplification assembly 2, where the barrier layer 5 is located between the conductive cover plate 4 and the chip substrate 6, and the barrier layer 5, the chip substrate 6, and the conductive cover plate 4 define a housing 10, which includes a storage chamber 20, an elution chamber 21, a liquid path chamber 22, and an amplification chamber 23, which are sequentially connected to each other, and the elution chamber 21 and the second valve chamber 104 are connected to each other via a first channel 24. Optionally, the number of wash chambers is at least one. The chip substrate 6 includes a base plate 60, an electrode array 7, and an insulating layer 61, where the electrode array 7 is located on the base plate 60, and the insulating layer 61 covers the electrode array 7 and is close to the conductive cover plate 4, and the electrode array 7 is provided corresponding to the housing 10.

[0067] The amplification assembly 2 further includes a first hydrophobic layer 26, a hydrophilic layer 28, and a second hydrophobic layer 27. The first hydrophobic layer 26 is located on a side of the insulating layer 61 closer to the conductive cover plate 4. The conductive cover plate 4 includes a liquid path region 42 and an amplification region 43, which are connected to each other. The liquid path region 42 corresponds to the liquid path chamber 22, and the amplification region 43 corresponds to the amplification chamber 23. The hydrophilic layer 28 is located on a side of the amplification region 43 closer to the chip substrate 6, and is disposed opposite the first hydrophobic layer 26. A second hydrophobic layer 27 is applied to the side of the liquid path region 42 closer to the chip substrate 6, and the hydrophilic layer 28 is adjacent to the second hydrophobic layer 27.

[0068] Because a hydrophobic layer is present around the amplification region 43, the sample reagent can be in stable contact with the hydrophilic layer 28 without being pushed away by the generated air bubbles. Also, because the side of the chip substrate 6 facing the amplification region 43 is a hydrophobic layer, the contact area of ​​the sample reagent with the hydrophobic layer is smaller than that of the hydrophilic layer 28, thereby reducing or even preventing the generation of air bubbles, and also allowing the generated air bubbles to be efficiently discharged.

[0069] This embodiment also provides a method for manufacturing a conductive cover plate 4, which can be used to manufacture the above-mentioned microfluidic chip 304. First, a first hydrophobic layer 26 is applied to the conductive cover plate 4, and the conductive cover plate 4 with the applied first hydrophobic layer 26 is attached to a fixture (not shown). The fixture exposes only the amplification region 43 of the conductive cover plate 4, and the first hydrophobic layer 26 on the surface of the amplification region 43 is then erased with an erasing tool such as a cotton swab. Optionally, to further improve the hydrophilicity of the ITO glass, a hydrophilic coating can be applied to the surface of the amplification region 43 from which the hydrophobic coating has been erased, or the surface of the amplification region 43 can be hydrophilized by laser, etching, or plasma.

[0070] A heating unit 62 is provided in the base plate 60, and the heating unit 62 includes a first heating wire (not shown) and a second heating wire (not shown), with the first heating wires provided directly below both the first valve chamber 101 and the second valve chamber 104, and the second heating wire being located directly below the amplification chamber 23. The insulating layer 61 may be a dielectric layer.

[0071] The barrier layer 5 includes a mixture of adhesive and plastic beads, and the density of the adhesive and the density of the plastic beads are close to each other, for example, the ratio of the density of the adhesive to the density of the plastic beads is 95% or more. Optionally, the barrier layer 5 includes a gasket (not shown) and an adhesive (not shown). The distance between the conductive cover plate 4 and the chip substrate 6 is equal to the diameter of the plastic beads.

[0072] 8 and 9, the conductive cover plate 4 may be ITO glass. The conductive cover plate 4 includes a plate 40 and a cover 41, the plate 40 having a cavity 400 open at both the bottom and the top, the bottom opening of the cavity 400 being located on the side of the plate 40 closer to the chip substrate 6 and the top opening of the cavity 400 being located on the side of the plate 40 opposite the chip substrate 6, the cover 41 covering the top opening of the cavity 400, and the elution chamber 21 including the cavity 400. Optionally, the cover 41 and the plate 40 are integrally molded.

[0073] The extraction cover plate 3 is provided with a first sample injection hole 30, a second sample injection hole 31, and a third sample injection hole 32. The first sample injection hole 30 communicates with the lysing chamber 100, the second sample injection hole 31 communicates with the first cleaning chamber 102, and the third sample injection hole 32 communicates with the second cleaning chamber 103. The cover 41 is provided with a fourth sample injection hole 410, which communicates with the elution chamber 21. The lysing chamber 100 is pre-stored with a lysing solution, the first cleaning chamber 102 and the second cleaning chamber 103 are pre-stored with a cleaning solution, and the elution chamber 21 is pre-stored with an elution solution. The chip substrate 6 is provided with heating wires (not shown) at all positions corresponding to the first valve chamber 101 and the second valve chamber 104, and is used to melt the paraffin in the first valve chamber 101 and the second valve chamber 104. The electrode array 7 is located directly below the elution chamber 21 , the liquid path chamber 22 , and the amplification chamber 23 .

[0074] A surfactant (not shown) is stored in advance in the storage chamber 20, and the hardened surfactant may be freeze-dried or baked and then stored in the storage chamber 20 in advance. The storage chamber 20 and the elution chamber 21 are connected via a second channel 25, and the liquid path chamber 22 includes a liquid storage cavity 220 and a liquid separation cavity 221 arranged in sequence in the direction from the liquid path chamber 22 to the amplification chamber 23, the liquid storage cavity 220 being adjacent to the elution chamber 21, and the liquid separation cavity 221 being adjacent to the amplification chamber 23, and in the thickness direction of the microfluidics chip 304, the thickness c of the cavity 400 is 0.6 mm to 2 mm, the distance b between the top wall of the first channel 24 and the upper surface of the chip substrate 6 is less than 7 mm and not less than 4 mm, the distance d between the top wall of the second channel 25 and the upper surface of the chip substrate 6 is less than 7 mm and not less than 4 mm, and the distance a between the top wall of the liquid storage cavity 220 and the upper surface of the chip substrate 6 is less than 7 mm and not less than 4 mm.

[0075] 2, 3, and 10, the electrode array 7 includes, in order, an elution electrode region 70, a first liquid storage electrode region 71, a second liquid storage electrode region 72, a liquid separation electrode region 73, a liquid fixation electrode region 74, and an amplification electrode region 75. In projection onto the chip substrate 6, the elution electrode region 70 is located within the elution chamber 21, and the first liquid storage electrode region 71, the second liquid storage electrode region 72, the liquid separation electrode region 73, and the liquid fixation electrode region 74 are located within the liquid path chamber 22. The amplification electrode region 75 is located within the amplification chamber 23. The elution electrode region 70 includes one elution electrode 700. The first liquid storage electrode region 71 includes one adjacent first transition electrode 710 and a first liquid storage electrode group 8, and when projected onto the chip substrate 6, the first transition electrode 710 spans the elution chamber 21 and the liquid storage cavity 220, and the first transition electrode 710 is located between the elution electrode region 70 and the first liquid storage electrode group 8, which is located within the liquid storage cavity 220. Optionally, the elution electrode region 70 includes one or more elution electrodes 700, and the multiple elution electrodes 700 are arranged in sequence in a direction from the elution chamber 21 to the liquid path chamber 22.

[0076] The first liquid storage electrode group 8 includes three first liquid storage electrodes 80, which are arranged in order from the elution chamber 21 to the liquid path chamber 22. The area of ​​the next first liquid storage electrode 80 is larger than that of the previous first liquid storage electrode 80, and the previous first liquid storage electrode 80 is closer to the first transition electrode 710 than the next first liquid storage electrode 80. Each first liquid storage electrode 80 includes a first side edge and a second side edge extending in the direction from the elution chamber 21 to the liquid path chamber 22. The first side edge and the second side edge are opposite each other, and the distance between the first side edge and the second side edge of the next first liquid storage electrode is larger than the distance between the first side edge and the second side edge of the previous first liquid storage electrode. The first side edges of all the first liquid storage electrodes are arranged on the same straight line, and the second side edges of all the first liquid storage electrodes are arranged on the same straight line. Optionally, the number of first liquid storage electrodes 80 includes two or more.

[0077] The first transition electrode 710 and the first liquid storage electrode 80 adjacent to the first transition electrode 710 have first arc-shaped portions 802 that fit together, with the openings of the first arc-shaped portions 802 facing the first transition electrode 710, and the two adjacent first liquid storage electrodes 80 have second arc-shaped portions 804 that fit together, with the openings of the second arc-shaped portions 804 facing the first transition electrode 710. Three first arc-shaped segments 803 are spaced apart within the first arc-shaped portion 802, with the openings of the first arc-shaped segments 803 facing opposite the first transition electrode 710, and three second arc-shaped segments 805 are provided within the second arc-shaped portion 804, with the openings of the second arc-shaped segments 805 facing opposite the first transition electrode 710. Optionally, the number of first arc-shaped segments 803 is at least one, and the number of second arc-shaped segments 805 is at least one.

[0078] 11 and 12, the liquid-fixed electrode region 74 includes a liquid-fixed electrode 740 and a liquid-fixed electrode 741, which are adjacent to each other, and the second liquid storage electrode region 72 includes a second liquid storage electrode 90, a second liquid storage electrode 91, and a second liquid storage electrode 92, which are adjacent to each other. Optionally, the number of liquid-fixed electrodes and second liquid storage electrodes is determined according to design needs. The liquid separation electrode region 73 includes a liquid separation section 730 and a liquid transport section 736, the liquid transport section 736 being located between the liquid separation section 730 and the second liquid storage electrode 92, the liquid separation section 730 including a first electrode 731 and a second electrode 732 arranged side by side in the width direction of the liquid separation section 730, the area of ​​the second electrode 732 being smaller than the area of ​​the first electrode 731, the width a and width b of the liquid separation electrode region 73 being both smaller than the width c of the liquid fixing electrode region 74, and the width e of the second liquid storage electrode region 72 being larger than the width c of the liquid fixing electrode region 74.

[0079] The width direction c of the liquid-fixed electrode region 74 and the width direction d of the liquid transfer section 736 are perpendicular to each other, the width direction b of the first end of the liquid separation section 730 is parallel to the width direction d of the liquid transfer section 736, the first end of the liquid separation section 730 is close to the liquid transfer section 736, the width direction a of the second end of the liquid separation section 730 is parallel to the width direction c of the liquid-fixed electrode region 74, the second end of the liquid separation section 730 is close to the liquid-fixed electrode region 74. The shape of the second electrode 732 includes an isosceles triangle, with the hypotenuse of the second electrode 732 adjacent to the first electrode 731, and the liquid transfer section 736 and the liquid-fixed electrode region 74 adjacent to the two adjacent sides of the second electrode 732, respectively. Optionally, the shape of the second electrode 732 may be triangular, angular, or the like.

[0080] In this embodiment, the adjacent second liquid storage electrodes 91 and 92 have first arc-shaped edges 900 that fit together, with the opening of the first arc-shaped edges 900 facing the liquid separation electrode region 73. The adjacent liquid fixed electrodes 740 and 741 have second arc-shaped edges 743 that fit together, with the opening of the second arc-shaped edges 743 facing the liquid separation electrode region 73. A third arc-shaped edge 901 is provided within the first arc-shaped edge 900, with the opening of the third arc-shaped edge 901 facing opposite the liquid separation electrode region 73, and a fourth arc-shaped edge 744 is provided within the second arc-shaped edge 743, with the opening of the fourth arc-shaped edge 744 facing opposite the liquid separation electrode region 73. Optionally, the number of third arcuate edges 901 may be one or more, and the number of fourth arcuate edges 744 may be one or more.

[0081] The second liquid storage electrode region 72 further includes two second transition electrodes 720, and the liquid transport section 736 includes a liquid transport electrode 737 and a liquid transport electrode 738 adjacent to each other in the longitudinal direction of the liquid transport section 736, and the liquid transport electrode 738 adjacent to the second liquid storage electrode is located between the two second transition electrodes 720, and each second transition electrode 720 is adjacent to the second liquid storage electrode 92 and the liquid transport electrode 738, respectively.

[0082] The liquid separation portion 730 includes a first tooth-shaped portion 733, a second tooth-shaped portion 734, and a third tooth-shaped portion 735, the first tooth-shaped portion 733 being located on an edge of the liquid separation portion 730 close to the liquid fixation electrode region 74 and engaging with the edge of the liquid fixation electrode region 74, the second tooth-shaped portion 734 being located on an edge of the liquid separation portion 730 close to the liquid transfer portion 736 and engaging with the edge of the liquid transfer portion 736, and the third tooth-shaped portion 735 being located on an edge of the second electrode 732 close to the first electrode 731 and engaging with the first electrode 731.

[0083] 13 to 16, the present embodiment also provides a method for controlling the electrode array 7 when separating the liquid of the sample droplet.

[0084] The second liquid storage electrode 92 is covered with the sample mother droplets, and at this time, the liquid transport section 736 , the liquid separation section 730 , and the liquid fixed electrode 740 are energized to move the sample mother droplets to the liquid fixed electrode 740 .

[0085] Next, current is applied to the liquid fixed electrode 740, the liquid fixed electrode 741, the second electrode 732, and the liquid transfer electrode, and the sample mother droplets are gradually moved to the liquid fixed electrode 741. Because current is not applied to the first electrode 731 but is applied to the second electrode 732, the sample mother droplets in the liquid separation section 730 decrease in number but do not cease.

[0086] Next, electricity is applied to the liquid fixed electrode 740, the liquid fixed electrode 741, the liquid transfer electrode 737, the liquid transfer electrode 738, the second liquid storage electrode 92, the second liquid storage electrode 91, and the liquid storage electrode 90, the power to the liquid separation unit 730 is turned off, and the size of the split sample sub-droplets is fixed by the liquid fixed electrode 741 and the liquid fixed electrode 740. When electricity is applied to the liquid transfer electrode 737, the liquid transfer electrode 738, the second liquid storage electrode 92, the second liquid storage electrode 91, and the second liquid storage electrode 90, the sample mother droplets tend to move in the direction of the larger electrodes, the droplets in the liquid separation unit 730 gradually decrease, and the sample sub-droplets are separated from the sample mother droplets.

[0087] 17, this embodiment also provides a microfluidics system including the above-mentioned microfluidics chip 304, which includes a control terminal 300, a magnetic attraction device 301, a fluorescence detection device 302, and a driving circuit 303. The control terminal 300 is electrically connected to the electrode array 7 via the driving circuit 303, and the control terminal 300 sends control commands to the driving circuit 303. The driving circuit 303 is electrically connected to the electrode array 7 and is used to control the current flow state of the electrode array 7. The control terminal 300 controls and moves the magnetic attraction device 301, thereby moving the test sample containing magnetic beads and reagents within the extraction assembly 1, and the fluorescence detection device 302 is used to detect the results of amplifying the test sample in the amplification region 43.

[0088] Referring to Figures 4 and 5, this embodiment also provides a method of using the above-mentioned microfluidics chip 304, in which a test sample (not shown) containing magnetic beads is controlled by a magnetic attraction device 301 (not shown) to move through the dissolution chamber 100, the washing chamber and into the elution chamber 21, and then the magnetic beads are controlled by the magnetic attraction device 301 to move back and forth between the storage chamber 20 and the elution chamber 21, thoroughly and uniformly mixing the surfactant and the test sample droplets, and then the electrode array 7 is energized so that the sample droplets enter the liquid storage cavity 220 from the elution chamber 21.

[0089] It should be noted that the above is merely a preferred embodiment of the present invention, but the design concept of the invention is not limited to this, and any insubstantial changes made to the present invention using this concept are all included in the scope of protection of the present invention. [Industrial Applicability]

[0090] In the microfluidic chip and method of use, microfluidic system, and method for manufacturing a conductive cover plate of the present invention, the surface coating of the conductive cover plate is modified to ensure stable sample reagent presence in the amplification region and facilitate observation while reducing the risk of bubbles occurring during the PCR process. The chip has a simple and compact structure, and by optimizing the size of specific cavities, it is possible to simplify the process and reduce costs. By optimizing the structure and configuration of the electrode array, it is possible to conveniently store eluent, achieve a larger capacity, improve detection efficiency, and ensure that sample droplets in the elution chamber are completely transported into the liquid channel chamber. Furthermore, during the liquid separation process, it is possible to control the uniform fragmentation of sample droplets and precisely control the fragmentation size of the sample droplets.

Claims

1. A microfluidics chip comprising an extraction assembly and an amplification assembly, the extraction assembly includes a lysis chamber, a first valve chamber, a wash chamber, and a second valve chamber, which are in sequential communication with each other; the amplification assembly includes a chip substrate, a barrier layer, and a conductive cover plate; the barrier layer is positioned between the chip substrate and a conductive cover plate, the barrier layer, the chip substrate, and the conductive cover plate define an elution chamber, a liquid path chamber, and an amplification chamber, which are sequentially in communication with each other, and the elution chamber and the second valve chamber are in communication with each other via a first channel; an electrode array is provided on the chip substrate; the conductive cover plate has a cavity with an open bottom on a side closer to the chip substrate, and the elution chamber includes the cavity; the electrode array is located directly below the elution chamber, the liquid path chamber, and the amplification chamber; the amplification assembly further includes a storage chamber for storing a surfactant, the storage chamber and the elution chamber communicating with each other via a second channel; the fluid path chamber includes a fluid storage cavity, the fluid storage cavity being adjacent to the elution chamber; A microfluidics chip, characterized in that the thickness of the cavity in the thickness direction of the microfluidics chip is 0.6 mm to 2 mm.

2. the electrode array includes an elution electrode region and a first liquid storage electrode region adjacent to each other, the elution electrode region including at least one elution electrode, the elution electrode being located within the elution chamber in projection onto the chip substrate; 2. The microfluidics chip of claim 1, wherein the first liquid storage electrode region includes one first transition electrode and a first liquid storage electrode group adjacent to each other, and when projected onto the chip substrate, the first transition electrode spans the elution chamber and the liquid storage cavity, and the first transition electrode is located between the elution electrode region and the first liquid storage electrode group, and the first liquid storage electrode group is located within the liquid storage cavity.

3. 3. The microfluidics chip of claim 2, wherein the first liquid storage electrode group includes at least two first liquid storage electrodes, the plurality of first liquid storage electrodes being arranged in sequence in a direction from the elution chamber to the liquid path chamber, the area of ​​the next first liquid storage electrode being larger than the area of ​​the previous first liquid storage electrode, and the previous first liquid storage electrode being closer to the first transition electrode than the next first liquid storage electrode.

4. 4. The microfluidics chip of claim 3, wherein each of the first liquid storage electrodes includes a first side edge and a second side edge extending in a direction from the elution chamber to the liquid path chamber, the first side edge and the second side edge being opposite each other, and the distance between the first side edge and the second side edge of a next first liquid storage electrode is greater than the distance between the first side edge and the second side edge of a previous first liquid storage electrode.

5. 5. The microfluidic chip according to claim 4, wherein the first side edges of all the first liquid storage electrodes are arranged on the same straight line, and the second side edges of all the first liquid storage electrodes are arranged on the same straight line.

6. the first transition electrode and the first liquid storage electrode adjacent to the first transition electrode have first arc-shaped portions that fit together, and an opening of the first arc-shaped portion faces the first transition electrode; The microfluidic chip according to any one of claims 3 to 5, characterized in that two adjacent first liquid storage electrodes have second arc-shaped portions that fit together, and openings of the second arc-shaped portions face the first transition electrode.

7. at least one first arcuate segment is provided within the first arcuate portion, an opening of the first arcuate segment facing the first transition electrode; The microfluidic chip of claim 6, wherein at least one second arcuate segment is provided within the second arcuate portion, and an opening of the second arcuate segment faces the first transition electrode.

8. the conductive cover plate includes a plate and a cover; The microfluidics chip according to any one of claims 1 to 5, characterized in that the cavity has an open top, and the cover is provided to cover the open top of the cavity.

9. an amplifier assembly, the amplifier assembly including a chip substrate, a barrier layer, and a conductive cover plate; the chip substrate is disposed opposite the conductive cover plate; the chip substrate includes a base plate, an electrode array, and an insulating layer, the electrode array being located on the base plate, and the insulating layer covering the electrode array and being close to the conductive cover plate; the barrier layer is positioned between the chip substrate and the conductive cover plate, a receiving portion is formed between the barrier layer, the chip substrate, and the conductive cover plate, and the electrode array corresponds to the receiving portion; the electrode array includes a second liquid storage electrode region, a liquid separation electrode region, and a liquid fixation electrode region, which are adjacent in sequence; A microfluidic chip, wherein both the width of the liquid fixing electrode region and the width of the second liquid storage electrode region are larger than the width of the liquid separation electrode region, the liquid-fixed electrode region includes at least one liquid-fixed electrode; the second liquid storage electrode region includes at least one second liquid storage electrode; the liquid separation electrode region includes a liquid separation section, the liquid separation section including a first electrode and a second electrode provided adjacent to each other in a width direction of the liquid separation section, A microfluidics chip, characterized in that the area of ​​the second electrode is smaller than the area of ​​the liquid-fixing electrode.

10. 10. The microfluidic chip of claim 9, wherein the liquid separation electrode region further includes a liquid transfer section, the liquid transfer section including at least one liquid transfer electrode, and the liquid transfer section is located between the liquid separation section and the second liquid storage electrode region.

11. The microfluidics chip of claim 10, wherein the width direction of the liquid-fixed electrode intersects with the width direction of the liquid transport section, the width direction of a first end of the liquid separation section is parallel to the width direction of the liquid transport section, the first end of the liquid separation section is close to the liquid transport section, the width direction of a second end of the liquid separation section is parallel to the width direction of the liquid-fixed electrode region, and the second end of the liquid separation section is close to the liquid-fixed electrode region.

12. The microfluidics chip according to claim 11, wherein the width direction of the liquid fixing electrode region and the width direction of the liquid transport section are perpendicular to each other.

13. the second liquid storage electrode region includes two or more second liquid storage electrodes adjacent to each other in order, and the two adjacent second liquid storage electrodes have first arc-shaped edges that fit together, and openings of the first arc-shaped edges face the liquid separation electrode region; The microfluidics chip according to any one of claims 9 to 12, characterized in that the liquid fixation electrode region includes two or more of the liquid fixation electrodes adjacent to each other in sequence, and the two adjacent liquid fixation electrodes have second arc-shaped edges that fit together, and the openings of the second arc-shaped edges face the liquid separation electrode region.

14. a third arcuate edge is provided within the first arcuate edge, the opening of the third arcuate edge being opposite the liquid separation electrode region; The microfluidic chip of claim 13, wherein a fourth arcuate edge is provided within the second arcuate edge, and an opening of the fourth arcuate edge faces the liquid separation electrode region.

15. 13. The microfluidics chip according to claim 10, wherein the area of ​​the first electrode is larger than the area of ​​the second electrode.

16. The microfluidics chip of claim 15, characterized in that the shape of the second electrode includes an isosceles triangle, the hypotenuse of the second electrode is adjacent to the first electrode, and the liquid transport section and the liquid fixing electrode region are each adjacent to two adjacent sides of the second electrode.

17. The microfluidics chip according to any one of claims 10 to 12, characterized in that the second liquid storage electrode region further includes two second transition electrodes, a liquid transport electrode adjacent to the second liquid storage electrode is located between the two second transition electrodes, and each of the second transition electrodes is adjacent to the second liquid storage electrode and the liquid transport electrode, respectively.

18. the liquid separating portion includes a first tooth portion, a second tooth portion, and a third tooth portion; the first tooth-shaped portion is located at an edge of the liquid separating portion close to the liquid fixed electrode region and engages with the edge of the liquid fixed electrode region; the second tooth-like portion is located at an edge of the liquid separating portion close to the liquid transferring portion and engages with the edge of the liquid transferring portion; The microfluidics chip according to any one of claims 10 to 12, characterized in that the third tooth-like portion is located on an edge of the second electrode close to the first electrode and is fitted with the first electrode.

19. an amplification assembly, the amplification assembly including a chip substrate, a conductive cover plate, a barrier layer, and a first hydrophobic layer; the chip substrate is disposed opposite the conductive cover plate; the chip substrate includes a base plate, an electrode array, and an insulating layer; the first hydrophobic layer is located on a side of the insulating layer that is closer to the conductive cover plate; the electrode array is located on the base plate, and the insulating layer covers the electrode array; the barrier layer is located between the chip substrate and the conductive cover plate, and a liquid path chamber and an amplification chamber are formed between the barrier layer, the chip substrate, and the conductive cover plate, the liquid path chamber, the amplification chamber, and the electrode array being provided correspondingly; the conductive cover plate includes a liquid path region and an amplification region connected to each other, the liquid path region corresponding to the liquid path chamber, and the amplification region corresponding to the amplification chamber; the amplification assembly further comprising an adjacent second hydrophobic layer and a hydrophilic layer; the hydrophilic layer is located on a side of the amplification region closer to the chip substrate; A microfluidics chip characterized in that the second hydrophobic layer is located on a side of the liquid path region closer to the chip substrate, and the second hydrophobic layer and the hydrophilic layer are arranged opposite the first hydrophobic layer.

20. 20. The microfluidic chip of claim 19, wherein the barrier layer comprises a mixture of adhesive and plastic beads, and the spacing between the chip substrate and the conductive cover plate is equal to the diameter of the plastic beads.

21. 21. The microfluidic chip of claim 20, wherein the ratio of the density of the adhesive to the density of the plastic beads is 95% or more.

22. the microfluidics chip includes an extraction assembly, the extraction assembly in communication with the amplification assembly; the extraction assembly includes a lysis chamber, a wash chamber, and an elution chamber, which are in sequential communication with each other; the lysis chamber, the washing chamber, and the elution chamber are separated by a paraffin valve; The microfluidics chip described in any one of claims 19 to 21, characterized in that the microfluidics chip further includes a heating unit provided in the chip substrate, the heating unit including a first heating wire and a second heating wire provided in the base plate, the first heating wire corresponding to the paraffin valve, and the second heating wire corresponding to the amplification chamber.

23. A method for manufacturing a conductive cover plate applied to the microfluidic chip according to any one of claims 19 to 22, comprising: a manufacturing method comprising the steps of applying a first hydrophobic layer on a conductive cover plate, attaching the conductive cover plate with the applied first hydrophobic layer to a fixture, and erasing the first hydrophobic layer on the surface of the amplification region with an erasing tool.

24. 24. The method for manufacturing a conductive cover plate according to claim 23, further comprising the step of: subjecting a surface of the amplification region from which the first hydrophobic layer has been removed to a hydrophilic treatment.

25. The method of claim 23 , wherein the fixture exposes only the amplification region of the conductive cover plate.

26. A method of using a microfluidics chip, comprising: The method is used for the microfluidic chip according to any one of claims 1 to 8, and the method comprises: A method of use, comprising the steps of: controlling a test sample droplet containing magnetic beads using a magnetic attraction device to move through the lysis chamber, the washing chamber in sequence, and into the elution chamber; controlling the magnetic beads using the magnetic attraction device to move back and forth between the storage chamber and the elution chamber; and powering on the electrode array so that the sample droplet in the elution chamber enters a liquid storage cavity.

27. 1. A microfluidics system comprising: The system includes the microfluidics chip according to any one of claims 1 to 22, and further includes a driving circuit and a control terminal; the control terminal is electrically connected to the driving circuit and is used to send control commands to the driving circuit; A microfluidics system, characterized in that the driving circuit is electrically connected to the electrode array and is used to control changes in the electrical conduction state of the electrode array.

28. The microfluidics system of claim 27, further comprising a magnetic attraction device and a fluorescence detection device, wherein the magnetic attraction device is used to control the sample to move within the extraction assembly, and the fluorescence detection device is used to detect the results of amplifying the sample within the amplification region.

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