A digital microfluidic chip for realizing multi-detection of allergens

Abstract

The invention relates to a digital microfluidic chip for realizing allergen multi-joint detection, comprising: an upper substrate is provided with a grounding electrode layer, the grounding electrode layer is connected to a first power supply; the grounding electrode layer is provided with a serum inlet and a plurality of detection reagent inlets that penetrate through its wall thickness, and the plurality of detection reagent inlet arrays are arranged around the serum inlet; a channel layer is provided with a through microchannel, and the microchannel is connected with the serum inlet and the detection reagent inlet; a plurality of terminal electrode arrays of the electrode array layer of the substrate are arranged around the central electrode, and the terminal electrode is connected to the central electrode through a sub-electrode group; the central electrode, the sub-electrode group and the terminal electrode are respectively connected to a second power supply; the upper substrate, the dielectric layer, the electrode array layer and the substrate are stacked in sequence from top to bottom; the serum inlet and the central electrode, the detection reagent inlet and the terminal electrode are arranged one by one up and down; the first power supply and the second power supply form a control circuit loop. The invention greatly improves detection efficiency and reduces costs.

Description

Technical Field

[0001] The invention relates to the technical field of allergen detection, and in particular to a digital microfluidic chip for realizing multiple allergen detection. Background Art

[0002] The World Allergy Organization has conducted an epidemiological survey on allergic diseases in 30 countries with a total population of 1.2 billion. The survey results show that nearly 22% of the population suffers from allergic diseases mediated by immunoglobulin E (IgE). Nowadays, allergic diseases have become the sixth largest chronic disease in the world, causing inconvenience in life and work for countless patients and bringing economic burden to the national health care system. In order to effectively treat allergic diseases and reduce the pain and economic pressure of patients caused by the disease and treatment, the cause of sensitization must be clarified first. Therefore, allergen detection is the core of allergic disease treatment.

[0003] There are two methods for allergen detection: in vivo detection and in vitro detection. In vivo detection is performed by observing the patient's reaction after the allergen comes into contact with the body, and mainly includes skin prick test, patch test, etc. In vivo detection is generally simple to operate and low in cost. In vitro detection is to draw blood or secretions from the patient for specific antibody detection. Compared with in vivo detection, since there is no need to allow allergens to enter the patient's body, in vitro detection has higher safety and is not affected by factors such as skin and drugs. It is suitable for various patient groups such as children, the elderly and those with hypersensitivity reactions, and has gradually become the preferred choice for allergen detection. However, in vitro testing often requires expensive instruments and equipment, and the number of tests is limited, the test speed is slow, and the test results often need to wait for several days or even longer.

[0004] Microfluidic chips can precisely manipulate microfluids in micron-scale microtubes, and can integrate sampling, dilution, sample addition, reaction, splitting, detection and other operation steps that need to be completed in the laboratory into reusable microfluidic chips. The on-chip droplet driving methods mainly include dielectric wetting drive, thermal capillary drive, light-induced drive, surface acoustic wave drive and magnetic drive. Among them, digital microfluidic technology (DMF) based on dielectric wetting effect (Electrowetting on Dielectric, EWOD) can drive a single or several droplets to move freely around on the chip plane, with the advantages of low reagent consumption, low energy consumption, fast reaction speed, high throughput, fast reaction, short detection time, and can realize multiple parallel biochemical reactions. At present, digital microfluidics has been applied to many biochemical medical directions including enzymatic reactions, immunoassays, protein analysis, tissue engineering and cell applications.

[0005] Digital microfluidics technology can make up for the time cost shortcomings of in vitro testing. In addition, the electrodes for electrochemical analysis and detection are small in size and easy to prepare. Once the technology matures, it can also reduce economic costs. It is one of the research directions with great development value in the field of medical biomarker detection.

[0006] However, the existing digital microfluidics technology has the problem of low efficiency in allergen detection. Summary of the invention

[0007] Therefore, the technical problem to be solved by the invention is to overcome the problem of low efficiency of allergen detection in the prior art.

[0008] In order to solve the above technical problems, on the one hand, the invention provides a digital microfluidic chip for realizing allergen multi-detection, comprising:

[0009] The upper substrate is provided with a grounding electrode layer, which is connected to the first power supply; the grounding electrode layer is provided with a serum inlet and a plurality of detection reagent inlets that penetrate through the thickness of the grounding electrode layer, and the plurality of detection reagent inlet arrays are arranged around the serum inlet;

[0010] The channel layer is provided with a through microchannel, and the microchannel is connected with the serum inlet and the detection reagent inlet;

[0011] Dielectric layer;

[0012] The substrate is provided with an electrode array layer, the electrode array layer includes a central electrode, a plurality of sub-electrode groups and a plurality of terminal electrodes; the plurality of terminal electrode arrays are arranged around the central electrode, the terminal electrodes are connected to the central electrode through the sub-electrode groups; the central electrode, the sub-electrode groups and the terminal electrodes are respectively connected to a second power supply;

[0013] Among them, the upper substrate, dielectric layer, electrode array layer and base are stacked in sequence from top to bottom; the serum inlet and the central electrode are arranged in correspondence with each other, and the detection reagent inlet and the end point electrode are arranged in correspondence with each other; the microchannel is located between the dielectric layer and the upper substrate; the first power supply and the second power supply form a control circuit loop.

[0014] In one embodiment of the invention, the electrode array layer further comprises at least one split electrode;

[0015] The split electrode is located between the central electrode and the terminal electrode, the split electrode is connected to the central electrode through a split electrode group, and the split electrode is connected to the terminal electrode through at least two split electrode groups.

[0016] In one embodiment of the invention, the sub-electrode group includes a plurality of driving electrodes, the plurality of driving electrodes are arranged in sequence, and two adjacent driving electrodes are insulated from each other.

[0017] In one embodiment of the invention, in the electrode group, two adjacent driving electrodes are provided with a connecting portion at the connection between the two electrodes; the connecting portion includes a slot and a protruding portion;

[0018] Of two adjacent driving electrodes, the protrusion of one is inserted into the groove of the other.

[0019] In one embodiment of the invention, the sub-electrode group and the central electrode are provided with a connecting portion at the connection between the two; the connecting portion includes a slot and a protrusion;

[0020] Of two adjacent driving electrodes, the protrusion of one is inserted into the groove of the other.

[0021] In one embodiment of the invention, the lengths of the plurality of sub-electrode groups are equal.

[0022] In one embodiment of the invention, the terminal electrodes are arranged in a regular polygonal array with the central electrode as the center; and the sub-electrode groups are arranged along straight lines.

[0023] In one embodiment of the invention, the microchannel includes a central hole, a plurality of guide grooves, and a plurality of reaction grooves;

[0024] A plurality of reaction slots are arranged in array around the central hole, and the reaction slots are connected with the central hole through guide slots;

[0025] The central hole and the serum inlet are arranged one by one in vertical correspondence, the guide grooves and the sub-electrode groups are arranged one by one in vertical correspondence, and the reaction grooves and the detection reagent inlets are arranged one by one in vertical correspondence.

[0026] In one embodiment of the invention, the present application further includes a first hydrophobic layer, and the first hydrophobic layer is stacked on the dielectric layer.

[0027] In one embodiment of the invention, the upper substrate further comprises a second hydrophobic layer, which is stacked below the ground electrode layer; the serum inlet and the detection reagent inlet respectively penetrate the second hydrophobic layer.

[0028] In one embodiment of the invention, the central electrode, the sub-electrode group and the terminal electrode are respectively connected to the second power supply through external leads, and a relay is provided on the external leads;

[0029] The control circuit loop includes a control unit, which is electrically connected to the relay and the second power supply.

[0030] In one embodiment of the invention, the base and the upper substrate are made of ITO conductive glass.

[0031] In one embodiment of the invention, the height of the microchannel is 200 um to 100 mm.

[0032] The above technical solution of the invention has the following advantages compared with the prior art:

[0033] The digital microfluidic chip for realizing multi-joint detection of allergens described in the present invention is provided with a ground electrode layer and an electrode array layer, an upper substrate and an electrode array layer, the upper substrate includes a ground electrode layer, the ground electrode layer is connected to a first power supply, the central electrode, the sub-electrode group and the terminal electrode of the electrode array layer are respectively connected to a second power supply, and the first power supply and the second power supply are connected to form a loop. In this way, an electric field can be formed between the ground electrode layer and the conductive central electrode, the ground electrode layer and the conductive sub-electrode group, or the ground electrode layer and the conductive terminal electrode. In addition, a dielectric layer and a microchannel are provided between the electrode array layer and the ground electrode layer, and the upper substrate is provided with a detection reagent inlet and a serum inlet, so that after the serum dripped from the serum inlet enters the microchannel, the central electrode, the terminal electrode and some of the sub-electrode groups in the multiple sub-electrode groups are controlled to be connected and connected with the second power supply, thereby realizing the splitting of the serum and transporting it to the corresponding position along the corresponding path, and then reacting with the detection reagent solution dripped from the detection reagent inlet. In this way, multiple detections can be realized by injecting a serum solution once, thereby greatly improving the detection efficiency and reducing the cost. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] In order to make the content of the invention more clearly understood, the invention is further described in detail below based on the specific embodiments of the invention and in conjunction with the accompanying drawings, wherein

[0035] Figure 1 It is a three-dimensional schematic diagram of a digital microfluidic chip for realizing multi-link detection of allergens in a preferred embodiment of the present invention;

[0036] Figure 2 yes Figure 1 A cross-sectional view of a digital microfluidic chip for realizing multi-link detection of allergens at position BB is shown;

[0037] Figure 3 yes Figure 1 A schematic diagram of an electrode array layer in a digital microfluidic chip for realizing multi-detection of allergens is shown;

[0038] Figure 4 yes Figure 3 An enlarged view of position C of an electrode array layer in a digital microfluidic chip for realizing multi-link detection of allergens is shown;

[0039] Figure 5 yes Figure 3 A schematic diagram of the arrangement of a central electrode, a plurality of sub-electrode groups and a plurality of end-point electrodes in an electrode array layer in a digital microfluidic chip for realizing multi-joint detection of allergens is shown;

[0040] Figure 6 yes Figure 3Another schematic diagram of the arrangement of a central electrode, a plurality of sub-electrode groups and a plurality of end-point electrodes in an electrode array layer in a digital microfluidic chip for realizing multi-joint detection of allergens is shown;

[0041] Figure 7 yes Figure 3 A schematic diagram of another arrangement of a central electrode, a plurality of sub-electrode groups and a plurality of end-point electrodes in an electrode array layer in a digital microfluidic chip for realizing multi-joint detection of allergens is shown;

[0042] Figure 8 yes Figure 3 A schematic diagram of the arrangement of microchannels in a digital microfluidic chip for realizing multi-joint detection of allergens (the corresponding schematic diagram of the arrangement of the central electrode, multiple sub-electrode groups and multiple end-point electrodes is shown in FIG. Figure 7 ).

[0043] Description of the Figures in the Specification:

[0044] 100, upper substrate; 110, ground electrode layer; 111, serum inlet; 112, detection reagent inlet; 120, second hydrophobic layer;

[0045] 200, channel layer; 210, microchannel; 211, central hole; 212, guide groove; 213, reaction groove; 220, spacer;

[0046] 300, dielectric layer;

[0047] 400, substrate; 410, electrode array layer; 411, central electrode; 412, sub-electrode group; 412a, driving electrode; 413, terminal electrode; 414, split electrode; 415, slot; 416, protrusion;

[0048] 500, a first hydrophobic layer;

[0049] 600, external lead. DETAILED DESCRIPTION

[0050] The invention is further described below in conjunction with the accompanying drawings and specific embodiments so that those skilled in the art can better understand the invention and implement it, but the embodiments are not intended to limit the invention.

[0051] Reference Figure 1 to Figure 6 As shown, on the one hand, the invention provides a digital microfluidic chip for realizing multi-joint detection of allergens, comprising:

[0052] The upper substrate 100 is provided with a grounding electrode layer 110, which is connected to a first power source; the grounding electrode layer 110 is provided with a serum inlet 111 and a plurality of detection reagent inlets 112 that penetrate through the thickness of the grounding electrode layer 110, and the plurality of detection reagent inlets 112 are arranged in an array around the serum inlet 111;

[0053] The channel layer 200 is provided with a through microchannel 210, and the microchannel 210 is connected with the serum inlet 111 and the detection reagent inlet 112;

[0054] Dielectric layer 300;

[0055] The substrate 400 is provided with an electrode array layer 410, and the electrode array layer 410 includes a central electrode 411, a plurality of sub-electrode groups 412, and a plurality of terminal electrodes 413; the plurality of terminal electrodes 413 are arrayed around the central electrode 411, and the terminal electrodes 413 are connected to the central electrode 411 through the sub-electrode groups 412; the central electrode 411, the sub-electrode groups 412, and the terminal electrodes 413 are respectively connected to a second power source;

[0056] Among them, the upper substrate 100, the dielectric layer 300, the electrode array layer 410 and the base 400 are stacked in order from top to bottom; the serum inlet 111 and the central electrode 411 are arranged in correspondence with each other, and the detection reagent inlet 112 and the end point electrode 413 are arranged in correspondence with each other; the microchannel 210 is located between the dielectric layer 300 and the upper substrate 100; the first power supply and the second power supply form a control circuit loop. For example, the first power supply is a positive power supply and the second power supply is a negative power supply. For another example, the first power supply is a negative power supply and the second power supply is a positive power supply.

[0057] In some embodiments, the channel layer 200 includes a spacer 220, which is fixed between the upper substrate 100 and the base 400, and the space enclosed by the spacer 220 is the microchannel 210. In this way, the upper substrate 100 and the base 400 are connected and fixed by the spacer 220 with a fixed height.

[0058] In some other embodiments, the spacer 220 may be disposed between the dielectric layer 300 and the upper substrate 100 .

[0059] Specifically, the present embodiment is provided with a ground electrode layer 110 and an electrode array layer 410, an upper substrate 100 and an electrode array layer 410, the upper substrate 100 includes a ground electrode layer 110, the ground electrode layer 110 is connected to a first power source, the central electrode 411, the sub-electrode group 412 and the terminal electrode 413 of the electrode array layer 410 are respectively connected to a second power source, and the first power source and the second power source are connected to form a loop. In this way, an electric field can be formed between the ground electrode layer 110 and the conductive central electrode 411, the ground electrode layer 110 and the conductive sub-electrode group 412, or the ground electrode layer 110 and the conductive terminal electrode 413. In addition, a dielectric layer 300 and a microchannel 210 are provided between the electrode array layer 410 and the ground electrode layer 110, and the upper substrate 100 is provided with a detection reagent inlet 112 and a serum inlet 111, so that after the serum dripped from the serum inlet 111 enters the microchannel 210, the central electrode 411, the terminal electrode 413 and some of the multiple sub-electrode groups 412 are connected to the second power supply by controlling the central electrode 411, the terminal electrode 413 and some of the multiple sub-electrode groups 412 to be connected and conducted, thereby achieving the splitting of the serum and transporting it to the corresponding position along the corresponding path, and then reacting with the detection reagent solution dripped from the detection reagent inlet 112. In this way, multiple tests can be achieved by injecting a serum solution once, thereby greatly improving the detection efficiency and reducing the cost.

[0060] Further, the electrode array layer 410 further includes at least one split electrode 414;

[0061] The split electrode 414 is located between the central electrode 411 and the terminal electrode 413 . The split electrode 414 is connected to the central electrode 411 through a split electrode group 412 . The split electrode 414 is connected to the terminal electrode 413 through at least two split electrode groups 412 .

[0062] Specifically, in this embodiment, a splitting electrode 414 is provided on the basis of the terminal electrode 413, and the splitting electrode 414 is connected to the terminal electrode 413 through the sub-electrode group 412, so that the serum solution after being split at the central electrode 411 can be transported to the splitting electrode 414, and then split again and run to the terminal electrode 413, so that multi-level splitting and transportation can be achieved, more joint detections can be achieved, and efficiency can be further improved. In this way, in the case of the same number of joint detections, the sub-electrode group 412 will not be densely distributed at the central electrode 411, thereby reducing manufacturing difficulty and reducing costs.

[0063] In some possible implementations, a plurality of splitting electrodes 414 are disposed between each central electrode 411 and each terminal electrode 413 of the electrode array layer 410 , thereby achieving multi-level splitting.

[0064] Furthermore, the sub-electrode group 412 includes a plurality of driving electrodes 412a, and the plurality of driving electrodes 412a are arranged in sequence, and two adjacent driving electrodes 412a are insulated from each other.

[0065] Specifically, the sub-electrode group 412 of this embodiment includes multiple driving electrodes 412a, so that the sub-electrode group 412 is spliced ​​by multiple driving electrodes 412a, so that sub-electrode groups 412 of different lengths can be made by splicing different numbers of driving electrodes 412a, which is more flexible and has a wider range of applications.

[0066] Furthermore, in the electrode group 412, two adjacent driving electrodes 412a are provided with a connecting portion at the connection between them; the connecting portion includes a slot 415 and a protrusion 416;

[0067] Among two adjacent driving electrodes 412 a , the protrusion 416 of one is inserted into the slot 415 of the other.

[0068] Specifically, in this embodiment, two adjacent driving electrodes 412a are provided with a connecting portion at the connection between the two, and the protrusion 416 of one of the two adjacent driving electrodes 412a is inserted into the card slot 415 of the other. In this way, the protrusion 416 and the card slot 415 provide a guiding function at the time gap between the two driving electrodes 412a when the serum solution moves from one driving electrode 412a to the other driving electrode 412a during the delivery process, so as to avoid the serum solution from being affected by insufficient driving force when moving between the two adjacent driving electrodes 412a.

[0069] Furthermore, the sub-electrode group 412 and the central electrode 411 are provided with a connecting portion at the connection between the two; the connecting portion includes a slot 415 and a protrusion 416;

[0070] Among two adjacent driving electrodes 412 a , the protrusion 416 of one is inserted into the slot 415 of the other.

[0071] Specifically, the effects achieved by this embodiment are the same as those of the above embodiment, which will not be described in detail here.

[0072] Furthermore, the lengths of the plurality of sub-electrode groups 412 are equal.

[0073] Specifically, the lengths of the multiple sub-electrode groups 412 of this embodiment are equal, so that the splitting and transportation of the serum solution are performed synchronously, further saving time and improving efficiency.

[0074] See also Figure 7 Furthermore, the terminal electrodes 413 are arranged in a regular polygonal array with the central electrode 411 as the center; and the sub-electrode groups 412 are arranged along a straight line.

[0075] Specifically, the terminal electrodes 413 of the present embodiment are arranged in a regular polygon array with the central electrode 411 as the center, so that the arrangement of the terminal electrodes 413 is more regular, which reduces the process difficulty and is more beautiful.

[0076] See also Figure 8 , further, the microchannel 210 includes a central hole 211, a plurality of guide grooves 212 and a plurality of reaction grooves 213;

[0077] A plurality of reaction grooves 213 are arranged in an array around the central hole 211, and the reaction grooves 213 are connected to the central hole 211 through the guide grooves 212;

[0078] The central hole 211 is disposed one by one with the serum inlet 111 , the guide groove 212 is disposed one by one with the sub-electrode group 412 , and the reaction groove 213 is disposed one by one with the detection reagent inlet 112 .

[0079] Specifically, the present embodiment is provided with a guide groove 212, so as to improve the guiding effect during the transportation process of the split serum solution, making the transportation smoother and further improving the efficiency.

[0080] Continue to see Figures 1 to 8 Furthermore, the present application also includes a first hydrophobic layer 500 , and the first hydrophobic layer 500 is stacked on the dielectric layer 300 .

[0081] Specifically, the present embodiment provides a first hydrophobic layer 500, which is used to increase the initial contact angle of serum droplets, prevent serum droplets from adhering to the dielectric layer 300, reduce driving damping, prevent droplets from adhering to the wall, and maintain smooth transportation.

[0082] Furthermore, the upper substrate 100 further includes a second hydrophobic layer 120 , which is stacked below the ground electrode layer 110 ; the serum inlet 111 and the detection reagent inlet 112 respectively penetrate the second hydrophobic layer 120 .

[0083] Specifically, the second hydrophobic layer 120 is provided in this embodiment, and the second hydrophobic layer 120 is used to increase the initial contact angle of the serum droplets, prevent the serum droplets from adhering to the dielectric layer 300, reduce the driving damping, prevent the droplets from adhering to the wall, and maintain smooth transportation.

[0084] Furthermore, the central electrode 411, the sub-electrode group 412 and the terminal electrode 413 are respectively connected to the second power supply through the external lead 600, and the external lead 600 is provided with a relay;

[0085] The control circuit loop includes a control unit, which is electrically connected to the relay and the second power supply.

[0086] Specifically, the present embodiment is provided with a control unit, through which the conduction or disconnection of the central electrode 411, the sub-electrode group 412 and the terminal electrode 413 can be adjusted, and the voltage of the second power supply can be adjusted.

[0087] Furthermore, the base 400 and the upper substrate 100 are made of ITO conductive glass material.

[0088] Specifically, the substrate 400 and the upper substrate 100 of this embodiment are made of ITO conductive glass. ITO conductive glass is a transparent material with strong conductivity and can be easily etched into electrode patterns. Therefore, by using ITO conductive glass as the material of the upper substrate 100 and the substrate 400, the movement of droplets can be observed in real time.

[0089] Furthermore, the height of the microchannel 210 is 200 um to 100 mm.

[0090] Specifically, the microchannel 210 is a space for splitting and transporting the serum solution. If the size of the microchannel 210 is too small, it is not convenient for splitting and transporting the serum solution. If the size of the microchannel 210 is too large, it affects the electric field between the electrode array layer 410 and the ground electrode layer 110, thereby affecting the driving force for splitting and transporting the serum solution.

[0091] In some embodiments, the upper substrate 100, the base 400, the dielectric layer 300 and the electrode array layer 410 are all square. Among them, the side length of the upper substrate 100 and the base 400 is 48mm (the shape and size of the ground electrode and the second hydrophobic layer 120 included in the upper substrate 100 are the same as those of the upper substrate 100). The side lengths of the dielectric layer 300, the first hydrophobic layer 500 and the electrode array layer 410 are all 40mm. The length of the driving electrode 412a is 2mm, and there is a gap of 0.1mm between two adjacent driving electrodes 412a; the length of the end point electrode 413 is 5mm. The width of the external lead 600 is 0.25mm. The apertures of the serum inlet 111 and the detection reagent inlet 112 are both 1mm.

[0092] It should be noted that when using this application, the sample preparation work must be done first, and then the experimental operation is carried out. (For the convenience of the following description, in each sub-electrode group 412, the driving electrode 412a adjacent to the central electrode 411 is called the first driving electrode, the driving electrode 412a adjacent to the terminal electrode 413 is called the third driving electrode, and the remaining driving electrodes 412a are called the second driving electrodes).

[0093] Sample preparation: (1) Before the experiment, inject Aquape l glass antifog agent into the microchannel 210 and soak for 2 minutes, then clean the channel with acetone. Then place it in a constant temperature drying oven at 80°C for 30 minutes for drying, thereby enhancing the hydrophobicity of the surface of the microchannel 210 and preventing the serum solution from infiltrating the surface of the microchannel 210;

[0094] (2) using plasma water to dilute the serum solution to reduce its viscosity, and transferring a certain amount of the diluted serum solution to the middle serum inlet 111;

[0095] (3) The antigen detection reagent to be detected is dispersed in the plasma water, and then transferred to the detection reagent inlet 112 using a pipette.

[0096] Experimental operation: The relay can be controlled to be on and off. First, the control unit of the control circuit loop controls the opening of the first driving electrode 412a, and adjusts the first driving electrode 412a to a suitable voltage. The serum solution dripping from the serum inlet 111 into the microchannel 210 is split into multiple sub-droplets (the number of sub-droplets is the same as the number of the first driving electrodes 412a), and then multiple sub-electrode groups 412 are opened. The steps for opening each sub-electrode group 412 are as follows: first, the driving electrode 412a connected to the central electrode 411 in the sub-electrode group 412 is opened, and then the next driving electrode 412a is opened and the driving electrode 412a is closed until the terminal electrode 413 is opened, so that the serum solution is classified into multiple first sub-liquids, and each first sub-liquid is gradually moved in the electric field formed by the opening and closing of the corresponding driving electrode 412a until the first sub-liquid moves to the terminal electrode 413, and then the terminal electrode 413 is closed. In this way, above the terminal electrode 413, the first sub-liquid and the detection reagent droplet are fused and the reagents are mixed.

[0097] In other embodiments, a splitting electrode 414 is provided, so that when the first sub-liquid moves to the end point electrode 413, it is split again into smaller second sub-droplets, and the second sub-droplets split twice are sequentially transported to their respective splitting electrodes 414 to fuse with corresponding detection reagent droplets, thereby realizing multi-detection detection of allergens.

[0098] It should be noted that when the present application is tested, the allergen detection reagent is placed in the detection reagent inlet 112, and the serum is placed in the serum inlet 111; the serum enters the microchannel, and is split and separated by the conduction of different electrodes in the electrode array layer embedded in the glass substrate, and transported to each allergen detection reagent for detection, thereby achieving simultaneous and efficient detection of multiple allergens.

[0099] It should be noted that the present application applies voltage to the electrode array layer 410 embedded under the dielectric layer 300 to change the wetting characteristics between the dielectric layer 300 and the conductive droplets (serum solution) attached to its surface, so that the liquid-solid contact angle changes, causing asymmetric deformation at both ends of the droplet, prompting a pressure difference to be generated inside the droplet, thereby achieving the operation and control of the droplet splitting or movement. The dielectric layer 300 is mainly used to accumulate charge so that the droplet can prevent electrolysis during the manipulation process.

[0100] On the other hand, the invention provides a method for preparing a digital microfluidic chip for realizing multi-detection of allergens, comprising:

[0101] On the substrate 400, a dry film photoresist is used to form an electrode array layer 410 by etching based on photolithography technology for controlling the movement of droplets;

[0102] A dielectric layer 300 is formed on the top of the electrode array layer 410 by using a vapor deposition method and a spin coating technique;

[0103] A serum inlet 111 and a plurality of detection reagent inlets 112 are formed by punching holes in the upper substrate 100;

[0104] The base 400 and the upper substrate 100 are aligned and bonded, and a microchannel 210 is formed between the dielectric layer 300 and the upper substrate 100 .

[0105] Obviously, the above embodiments are merely examples for the purpose of clear explanation and are not intended to limit the implementation methods. For those skilled in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the implementation methods here. The obvious changes or modifications derived from these are still within the scope of protection of the invention.

Claims

1. A digital microfluidic chip for realizing multi-detection of allergens. Features: include: The upper substrate is provided with a grounding electrode layer, and the grounding electrode layer is connected to the first power supply; the grounding electrode layer is provided with a serum inlet and a plurality of detection reagent inlets penetrating the thickness of the grounding electrode layer, and the plurality of detection reagent inlets are arrayed around the serum inlet; A microchannel, provided with a through microchannel, wherein the microchannel is connected to the serum inlet and the detection reagent inlet; Dielectric layer; The substrate is provided with an electrode array layer, the electrode array layer includes a central electrode, a plurality of sub-electrode groups and a plurality of terminal electrodes; the plurality of terminal electrode arrays are arranged around the central electrode, the terminal electrodes are connected to the central electrode through the sub-electrode groups; the central electrode, the sub-electrode groups and the terminal electrodes are respectively connected to a second power supply; Among them, the upper substrate, the dielectric layer, the electrode array layer and the base are stacked in sequence from top to bottom; the serum inlet and the central electrode are arranged in correspondence with each other, and the detection reagent inlet and the end point electrode are arranged in correspondence with each other; the microchannel is located between the dielectric layer and the upper substrate; the first power supply and the second power supply form a control circuit loop.

2. The digital microfluidic chip for realizing multi-link detection of allergens according to claim 1, Features: The electrode array layer further comprises at least one split electrode; The split electrode is located between the central electrode and the terminal electrode, the split electrode is connected to the central electrode through the split electrode group; and the split electrode is connected to the terminal electrode through at least two of the split electrode groups.

3. The digital microfluidic chip for realizing multi-link detection of allergens according to claim 1 or 2, Features: The sub-electrode group includes a plurality of driving electrodes, which are arranged in sequence, and two adjacent driving electrodes are insulated from each other.

4. The digital microfluidic chip for realizing multi-link detection of allergens according to claim 3, Features: In the sub-electrode group, two adjacent driving electrodes are provided with a connecting portion at the connection between the two; the connecting portion includes a slot and a protruding portion; The protrusion of one of the two adjacent driving electrodes is inserted into the slot of the other.

5. The digital microfluidic chip for realizing multi-link detection of allergens according to claim 4, Features: The sub-electrode group and the central electrode are both provided with a connecting portion at the connection between the two; the connecting portion includes a slot and a protrusion; The protrusion of one of the two adjacent driving electrodes is inserted into the slot of the other.

6. The digital microfluidic chip for realizing multi-link detection of allergens according to claim 1, Features: The lengths of the plurality of sub-electrode groups are equal.

7. The digital microfluidic chip for realizing multi-link detection of allergens according to claim 6, Features: The terminal electrodes are arranged in a regular polygonal array with the central electrode as the center; and the sub-electrode groups are arranged along a straight line.

8. The digital microfluidic chip for realizing multi-link detection of allergens according to claim 1, Features: The microchannel includes a central hole, a plurality of guide grooves and a plurality of reaction grooves; The plurality of reaction slot arrays are arranged around the central hole, and the reaction slots are connected to the central hole through the guide slots; The central hole is arranged one by one with the serum inlet, the guide groove is arranged one by one with the sub-electrode group, and the reaction groove is arranged one by one with the detection reagent inlet.

9. The digital microfluidic chip for realizing multi-link detection of allergens according to claim 1, Features: Also comprising a first hydrophobic layer, the first hydrophobic layer stacked on the dielectric layer; And / or, the upper substrate further includes a second hydrophobic layer, which is stacked below the ground electrode layer; and the serum inlet and the detection reagent inlet respectively penetrate the second hydrophobic layer.

10. The digital microfluidic chip for realizing multi-link detection of allergens according to claim 1, Features: The central electrode, the sub-electrode group and the terminal electrode are respectively connected to a second power supply through external leads, and a relay is provided on the external leads; The control circuit loop includes a control unit, and the control unit is electrically connected to the relay and the second power supply.

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