A reagent pre-embedding and sample injection device and method, and a digital microfluidic chip including the same

Abstract

The present invention provides a reagent pre-embedding and sample injection device and method, and a digital microfluidic chip including the same. The reagent pre-embedding and sample injection device includes a sample injection seat with a cavity structure, a reagent pack is placed in the top open end of the sample injection seat, the bottom liquid injection port of the sample injection seat is connected to the gap cavity of the digital microfluidic chip, and a needle puncture component is arranged below the reagent pack. When injecting samples, the reagent pack is squeezed, and the needle puncture conductive component punctures the reagent pack, and the reagent in the reagent pack flows into the reaction cavity of the digital microfluidic chip from the liquid injection port at the bottom of the sample injection seat. The reagent pre-embedding and sample injection device provided by the present invention is mainly used for digital microfluidic chips, and the reagents and other substances required for detection are quantitatively sealed in the reagent pack in advance, and the reagent pack is pre-embedded in the sample injection seat in advance and sealed together with the digital microfluidic chip. No manual operation is required by the user during the sample injection process, which can effectively prevent the inconvenience and waste caused by human operation errors.

Description

Technical Field

[0001] The invention belongs to the technical field of microfluidic chips, and relates to a reagent pre-embedding and sample injection device and method, and a digital microfluidic chip comprising the same. Background Art

[0002] Microfluidic chips integrate basic operating units such as sample preparation, reaction, separation, and detection in biological, chemical, and medical analysis processes into a chip with a microscale structure. The chip uses the principle of electrowetting technology to regulate the surface energy of solids and liquids through electric potential, and uses the imbalanced state of surface energy to drive the movement of liquids, thereby achieving precise control of microfluids. It can integrate the operating processes often required in the fields of biology, chemistry, and medicine, such as sampling, dilution, adding reagents, reaction, separation, and detection, on a digital microfluidic chip. Compared with traditional control methods, this technology can achieve less sample consumption, and at the same time has the advantages of high sensitivity, high precision, high throughput, and high integration. It can quickly realize the full process of biochemical reactions with low cost. The whole process reaction is fully enclosed without cross contamination and can be operated with one button, which greatly liberates the hands of operators.

[0003] During the liquid injection process of the microfluidic chip, the operator is usually required to use a pipette to absorb a certain amount of liquid sample, aim at the injection port, and inject the liquid completely into the reaction chamber. However, using a pipette to inject samples increases the cost of use and has high requirements on the operator's operating accuracy.

[0004] CN209406357U discloses a microfluidic chip for convenient liquid injection, including a substrate and a cover plate, wherein a plurality of microfluidic channels are arranged on the substrate, the substrate and the cover plate are bonded to form a whole, the microfluidic channels are located between the substrate and the cover plate, and the microfluidic channels also include a guide tube, wherein at least one guide hole is arranged on the cover plate, the guide hole is connected to the microfluidic channel, and one end of the guide tube is detachably connected to the guide hole.

[0005] CN204583216U discloses a microfluidic chip with autonomous microfluid movement, including a chip substrate and a cover plate. A microchannel with a V-groove cross-section is arranged on the chip substrate. The inlet depth of the microchannel is 10 to 800 microns, and the outlet depth of the channel is 20 to 800 microns. At the same time, the depth of the microchannel gradually deepens from the inlet to the outlet, and the change rule is ΔH=ΔLtanβ, ΔH is the channel depth increment, ΔL is the channel length increment, and 0<β<10 degrees.

[0006] CN110201614A discloses a flow chemical reaction device, comprising at least one reactor, wherein the reactor is provided with a sample outlet and at least two sample inlets; a sealed micro-channel is arranged in the reactor, and the at least two sample inlets are connected to the sample outlet through the micro-channel; each sample inlet is connected to an injection device through a channel, and the injection device is used to inject the sample into the reactor.

[0007] Traditional technologies use fully manually operated reaction plates, such as 96-well plates and 384-well plates; or continuous microfluidic devices and droplet microfluidics with syringe pumps; reaction plates have poor scalability and are usually fixed to 96 or 384 wells. Once the reaction is started, it is impossible to add new reaction groups midway. In practical applications, there are great limitations, and fully manual operation is time-consuming and labor-intensive, with low precision and prone to errors. The operation of microfluidic devices and droplet microfluidics is highly dependent on syringe pumps and is costly.

[0008] In addition, traditional technology usually requires the use of a pipette or an external mechanical pump, which has high manufacturing costs, complex operating procedures, high repeatability, and limited use environment. It is impossible to pre-embed the detection reagents on the digital microfluidic chip, and the liquid injection process is cumbersome and prone to waste and misoperation. Summary of the invention

[0009] In view of the deficiencies in the prior art, the purpose of the present invention is to provide a reagent pre-embedding and sample injection device and method, and a digital microfluidic chip including the same. The reagent pre-embedding and sample injection device provided by the present invention is mainly used for digital microfluidic chips. The reagents and other substances (related liquids, solids, or solid-liquid mixtures, etc.) required for detection are pre-quantitated and sealed in a reagent package, and the reagent package is pre-embedded in the sample injection seat in advance and sealed together with the digital microfluidic chip. During the sample injection process, no manual operation by the user is required, which can effectively prevent the inconvenience and waste caused by human operating errors.

[0010] To achieve this object, the present invention adopts the following technical solutions:

[0011] In the first aspect, the present invention provides a reagent pre-embedding and sample injection device, which comprises a sample injection seat with a cavity structure, a reagent package is placed in the top open end of the sample injection seat, a bottom liquid injection port of the sample injection seat is connected to the gap cavity of a digital microfluidic chip, and a puncture component is arranged under the reagent package. During sample injection, the reagent package is squeezed, the puncture conductive component punctures the reagent package, and the reagent in the reagent package flows into the reaction cavity of the digital microfluidic chip from the liquid injection port at the bottom of the sample injection seat.

[0012] The reagent pre-embedding and sample injection device provided by the present invention is mainly used for digital microfluidic chips. The reagents and other substances (related liquids, solids, or solid-liquid mixtures, etc.) required for detection are pre-quantitatively sealed in reagent packages, and the reagent packages are pre-embedded in the sample injection seat in advance and sealed together with the digital microfluidic chip. During the sample injection process, no manual operation by the user is required, which can effectively prevent the inconvenience and waste caused by human operating errors.

[0013] It should be noted that those skilled in the art can pre-embed an oil bag on the digital microfluidic chip and perform automatic oil filling according to the above design principles.

[0014] In addition, the present invention does not make specific requirements on the structural shape of the puncture component. It can be a separate sharp thorn structure or other structures that can puncture the reagent package. The puncture component can be fixed to the sample injection seat through other connecting parts or form an integrated structure with the sample injection seat.

[0015] As a preferred technical solution of the present invention, a sealing film is provided at the top opening of the sample injection seat, and the sealing film is in close contact with the reagent package.

[0016] Preferably, the sealing film is fixed to the top opening of the sample injection seat by hot melting or adhesive bonding.

[0017] Preferably, the sealing film is vacuum formed so that the sealing film adheres to the surface of the reagent package.

[0018] In the present invention, the sample injection seat is assembled on the digital microfluidic chip, the quantitatively packaged reagent package is placed in the sample injection seat, and the sealing film is fixed to the top opening of the sample injection seat by hot melting or adhesive bonding, and the fixed sealing film is attached to the surface of the reagent package by a vacuum blister process to ensure that a micro-vacuum environment is formed in the sample injection seat. The sealing film may be elastic or inelastic. Before pressing down, the reagent package is in a drum shape, and the sealing film also bulges accordingly. After pressing down, the reagent package is punctured, and the sealing film is pressed and depressed.

[0019] As a preferred technical solution of the present invention, the sample injection seat is divided into a receiving section and a puncture section, the reagent package is placed in the receiving section, and the puncture section is provided with a puncture component.

[0020] Preferably, the accommodating section and the needling section are integrally formed.

[0021] Preferably, the outer diameter of the accommodating section is larger than the outer diameter of the acupuncture section, the inner diameter of the accommodating section is larger than the inner diameter of the acupuncture section, and the joint between the accommodating section and the acupuncture section forms an annular accommodating platform, and the reagent package is placed on the accommodating platform.

[0022] Preferably, the acupuncture component is fixed to the inner wall of the acupuncture section via a support rod.

[0023] As a preferred technical solution of the present invention, the bottom injection port of the sample injection seat is connected to the gap cavity of the digital microfluidic chip through an injection tube.

[0024] As a preferred technical solution of the present invention, the reagent pre-embedding and sample injection device further comprises a pressing device, and the pressing device is located above the sample injection seat. During the sample injection process, the pressing device continuously squeezes the reagent package.

[0025] In the present invention, automatic liquid injection is achieved through a downward pressure device, which automatically moves downward to squeeze the reagent package. When the reagent package contacts the puncture component, the reagent package is punctured by the puncture component, and the reagent flows out and is injected into the gap cavity of the microfluidic chip through the injection tube. The entire injection process does not require manual operation by the user, which greatly improves the detection efficiency and reduces the detection cost.

[0026] It should be noted that the pressing device presses on the sealing film, and the sealing film has good elasticity and will not break after being deformed by pressing. If the sealing film rebounds after the pressing device rises, the pressing device needs to press down on the sealing film throughout the liquid injection stage. If the sealing film does not rebound after the pressing device rises and maintains a concave shape, the pressing device can rise and reset after completing one pressing.

[0027] As a preferred technical solution of the present invention, the volume of the reagent package is 100-300 μL, for example, it can be 100 μL, 120 μL, 140 μL, 160 μL, 180 μL, 200 μL, 220 μL, 240 μL, 260 μL, 280 μL or 300 μL, but is not limited to the listed values, and other unlisted values ​​within the numerical range are also applicable.

[0028] The reagent pre-embedding and sample injection device provided by the present invention can realize the liquid storage requirements of different systems within a certain system range by adjusting the size of the reagent package.

[0029] In a second aspect, the present invention provides a reagent pre-embedding and sample injection method, using the reagent pre-embedding and sample injection device described in the first aspect to inject a reagent into a gap cavity of a digital microfluidic chip, the method comprising:

[0030] The reagent pack is squeezed, and the needle-piercing conductive component pierces the reagent pack, and the reagent in the reagent pack flows into the reaction chamber of the digital microfluidic chip from the injection port at the bottom of the sample injection seat.

[0031] As a preferred technical solution of the present invention, the reagent pack is automatically squeezed by a pressing device.

[0032] In a third aspect, the present invention provides a digital microfluidic chip comprising the reagent pre-embedding and sample injection device described in the first aspect, the digital microfluidic chip comprising an electrode array, the surface of the electrode array is provided with the reagent pre-embedding and sample injection device described in the first aspect, the surface of the electrode array is also stacked with a dielectric layer and a hydrophobic layer in sequence, a transparent conductive cover is covered on the hydrophobic layer, a gap cavity is formed between the transparent conductive cover and the hydrophobic layer, and the bottom liquid injection port of the injection seat is connected to the gap cavity.

[0033] As a preferred technical solution of the present invention, the reagent pre-embedding and sample injection device is fixed to the edge of the electrode array;

[0034] Preferably, at least two reagent pre-embedding and sample injection devices are fixed side by side along the same edge of the electrode array;

[0035] Preferably, the reagents stored in each reagent pack are the same or different.

[0036] Compared with the prior art, the present invention has the following beneficial effects:

[0037] The reagent pre-embedding and sample injection device provided by the present invention is mainly used for digital microfluidic chips. The reagents and other substances (related liquids, solids, or solid-liquid mixtures, etc.) required for detection are quantitatively sealed in the reagent package in advance, and the reagent package is pre-embedded in the sample injection seat in advance and sealed together with the digital microfluidic chip. During the sample injection process, no manual operation by the user is required, which can effectively prevent the inconvenience and waste caused by human operation errors. . BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Figure 1 A schematic diagram of the structure of a digital microfluidic chip provided in a specific embodiment of the present invention;

[0039] Figure 2 A schematic diagram of the structure of a reagent pre-embedding and sample injection device provided in a specific embodiment of the present invention;

[0040] Figure 3 A three-dimensional diagram of a digital microfluidic chip provided in accordance with a specific embodiment of the present invention.

[0041] Among them, 1-sealing film; 2-reagent package; 3-sample injection seat; 4-puncture component; 5-liquid injection tube; 6-gap cavity; 7-transparent conductive cover; 8-hydrophobic layer; 9-dielectric layer; 10-electrode array; 11-accommodating section; 12-puncture section. DETAILED DESCRIPTION

[0042] It should be understood that, in the description of the present invention, the terms "center", "longitudinal", "lateral", "up", "down", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside", etc., indicating the orientation or positional relationship are based on the orientation or positional relationship shown in the drawings, and are only for the convenience of describing the present invention and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as a limitation on the present invention. In addition, the terms "first", "second", etc. are only used for descriptive purposes, and cannot be understood as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Therefore, the features defined as "first", "second", etc. may explicitly or implicitly include one or more of the features. In the description of the present invention, unless otherwise specified, "multiple" means two or more.

[0043] It should be noted that in the description of the present invention, unless otherwise clearly specified and limited, the terms "disposed", "connected", and "connected" should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection, or it can be indirectly connected through an intermediate medium, or it can be the internal communication of two elements. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood by specific circumstances.

[0044] The technical solution of the present invention is further described below with reference to the accompanying drawings and through specific implementation methods.

[0045] In a specific embodiment, the present invention provides a reagent pre-embedding and sample injection device, the reagent pre-embedding and sample injection device is as follows Figure 1 and Figure 2 As shown, it includes a sample injection seat 3 with a cavity structure, a reagent pack 2 is placed in the top open end of the sample injection seat 3, a bottom liquid injection port of the sample injection seat 3 is connected with the gap cavity 6 of the digital microfluidic chip, and a puncture component 4 is arranged under the reagent pack 2. When injecting samples, the reagent pack 2 is squeezed, the puncture conductive component punctures the reagent pack 2, and the reagent in the reagent pack 2 flows into the reaction cavity of the digital microfluidic chip from the liquid injection port at the bottom of the sample injection seat 3.

[0046] The reagent pre-embedding and injection device provided in this specific embodiment is mainly used for digital microfluidic chips. The reagents and other substances (related liquids, solids, or solid-liquid mixtures, etc.) required for detection are pre-quantitated and sealed in the reagent package 2, and the reagent package 2 is pre-embedded in the injection seat 3 in advance and sealed together with the digital microfluidic chip. During the injection process, no manual operation by the user is required, which can effectively prevent the inconvenience and waste caused by human operating errors.

[0047] It is understandable that, as an extension of this specific embodiment, those skilled in the art can pre-embed the oil bag on the digital microfluidic chip and automatically inject the oil according to the above design principles. In addition, there is no specific requirement for the structural shape of the puncture component 4, which can be a separate spike structure or other structures that can puncture the reagent package 2. The puncture component 4 can be fixed to the sample injection seat 3 through other connectors or form an integrated structure with the sample injection seat 3.

[0048] A sealing film 1 is provided at the top opening of the injection seat 3, and the sealing film 1 is fixed to the top opening of the injection seat 3 by hot melting or adhesive, and the sealing film 1 is vacuum-formed so that the sealing film 1 is attached to the surface of the reagent package 2. The injection seat 3 is assembled on a digital microfluidic chip, and the quantitatively packaged reagent package 2 is placed in the injection seat 3. The sealing film 1 is fixed to the top opening of the injection seat 3 by hot melting or adhesive, and the fixed sealing film 1 is attached to the surface of the reagent package 2 by vacuum forming process, so as to ensure that a micro-vacuum environment is formed in the injection seat 3. The sealing film 1 may be elastic or inelastic. Before pressing down, the reagent package 2 is in a drum shape, and the sealing film 1 also bulges accordingly. After pressing down, the reagent package 2 is punctured, and the sealing film 1 is pressed and depressed.

[0049] The sample injection seat 3 is divided into a receiving section 11 and a puncture section 12. The reagent pack 2 is placed in the receiving section 11, and the puncture section 12 is provided with a puncture component 4. The receiving section 11 and the puncture section 12 are integrally formed. The outer diameter of the receiving section 11 is larger than the outer diameter of the puncture section 12, and the inner diameter of the receiving section 11 is larger than the inner diameter of the puncture section 12. The docking point of the receiving section 11 and the puncture section 12 forms an annular receiving platform, the reagent pack 2 is placed on the receiving platform, the puncture component 4 is fixed to the inner wall of the puncture section 12 by a support rod, and the bottom injection port of the sample injection seat 3 is connected to the gap cavity 6 of the digital microfluidic chip through the injection tube 5. In this specific embodiment, automatic liquid injection is achieved through a downward pressure device, which automatically moves downward to squeeze the reagent package 2. When the reagent package 2 contacts the puncture component 4, the reagent package 2 is punctured by the puncture component 4, and the reagent flows out and is injected into the gap cavity 6 of the microfluidic chip through the injection tube 5. The entire injection process does not require manual operation by the user, which greatly improves the detection efficiency and reduces the detection cost.

[0050] It should be noted that the pressing device presses on the sealing film 1, and the sealing film 1 has good elasticity and will not break after being deformed by pressing. If the sealing film 1 rebounds after the pressing device rises, the pressing device needs to press down the sealing film 1 throughout the liquid injection stage. If the sealing film 1 does not rebound after the pressing device rises and maintains a concave shape, the pressing device can rise and reset after completing one pressing.

[0051] The reagent pre-embedding and sample injection device also includes a pressing device, which is located above the sample injection seat 3. During the sample injection process, the pressing device continuously squeezes the reagent package 2, and the volume of the reagent package 2 is 100 to 300 μL. The reagent pre-embedding and sample injection device provided in this specific embodiment can achieve the liquid storage requirements of different systems within a certain system range by adjusting the size of the reagent package 2.

[0052] In another specific embodiment, the present invention provides a reagent pre-embedding and sample injection method, the method comprising:

[0053] The reagent pack 2 is automatically squeezed by the pressing device, and the needle-piercing conductive component pierces the reagent pack 2, and the reagent in the reagent pack 2 flows into the reaction chamber of the digital microfluidic chip from the injection port at the bottom of the sample injection seat 3.

[0054] In another specific embodiment, the present invention provides a digital microfluidic chip, wherein the digital microfluidic chip is Figure 1 and Figure 3 As shown, it includes an electrode array 10, and a reagent pre-embedded and sample injection device is provided on the surface of the electrode array 10. Further, the reagent pre-embedded and sample injection device is fixed to the edge of the electrode array 10. Further, as shown in FIG. Figure 3 As shown, at least two reagent pre-embedding and sample injection devices are fixed side by side along the same edge of the electrode array 10, and the reagents stored in each reagent package 2 are the same or different.

[0055] The surface of the electrode array 10 is also stacked with a dielectric layer 9 and a hydrophobic layer 8 in sequence, a transparent conductive cover 7 is covered on the hydrophobic layer 8, a gap cavity 6 is formed between the transparent conductive cover 7 and the hydrophobic layer 8, and the bottom liquid injection port of the sample injection seat 3 is connected to the gap cavity 6.

[0056] The applicant declares that the above is only a specific implementation mode of the present invention, but the protection scope of the present invention is not limited thereto. Those skilled in the art should understand that any changes or substitutions that can be easily thought of by those skilled in the art within the technical scope disclosed by the present invention shall fall within the protection scope and disclosure scope of the present invention.

Claims

1. A reagent pre-embedding and sample injection device, characterized in that: The reagent pre-embedding and injection device comprises an injection seat with a cavity structure, a reagent package is placed in the top open end of the injection seat, the injection seat is divided into a receiving section and a puncture section, the outer diameter of the receiving section is larger than the outer diameter of the puncture section, the inner diameter of the receiving section is larger than the inner diameter of the puncture section, and the docking point of the receiving section and the puncture section forms an annular receiving platform, and the reagent package is placed on the receiving platform; the bottom injection port of the injection seat is connected to the gap cavity of the digital microfluidic chip, and a puncture component is arranged under the reagent package. When injecting the sample, the reagent package is squeezed, the puncture conduction component punctures the reagent package, and the reagent in the reagent package flows into the reaction cavity of the digital microfluidic chip from the injection port at the bottom of the injection seat.

2. The reagent pre-embedding and sample injection device according to claim 1, characterized in that: A sealing film is arranged at the top opening of the sample injection seat, and the sealing film is closely attached to the reagent package.

3. The reagent pre-embedding and sample injection device according to claim 2, characterized in that: The sealing film is fixed to the top opening of the sample injection seat by hot melting or adhesive sticking.

4. The reagent pre-embedding and sample injection device according to claim 2, characterized in that: The sealing film is vacuum-formed so that the sealing film adheres to the surface of the reagent package.

5. The reagent pre-embedding and sample injection device according to claim 1, characterized in that: The reagent pack is placed in the containing section, and the acupuncture section is provided with acupuncture components.

6. The reagent pre-embedding and sample injection device according to claim 1, characterized in that: The accommodating section and the acupuncture section are integrally formed.

7. The reagent pre-embedding and sample injection device according to claim 1, characterized in that: The acupuncture component is fixed to the inner wall of the acupuncture section through a supporting rod.

8. The reagent pre-embedding and sample injection device according to claim 1, characterized in that: The bottom injection port of the sample injection seat is connected to the gap cavity of the digital microfluidic chip through the injection tube.

9. The reagent pre-embedding and sample injection device according to claim 1, characterized in that: The reagent pre-embedding and sample injection device also includes a pressing device, which is located above the sample injection seat. During the sample injection process, the pressing device continuously squeezes the reagent package.

10. The reagent pre-embedding and sample injection device according to claim 1, characterized in that: The volume of the reagent pack is 100~300μL.

11. A reagent pre-embedding and sample injection method, characterized in that: The reagent pre-embedding and sample injection device according to any one of claims 1 to 10 is used to inject a reagent into the gap cavity of a digital microfluidic chip, the method comprising: The reagent pack is squeezed, and the needle-piercing conductive component pierces the reagent pack, and the reagent in the reagent pack flows into the reaction chamber of the digital microfluidic chip from the injection port at the bottom of the sample injection seat.

12. The reagent pre-embedding and sample injection method according to claim 11, characterized in that: The reagent pack is automatically squeezed by the pressing device.

13. A digital microfluidic chip comprising the reagent pre-embedding and sample injection device according to any one of claims 1 to 10, characterized in that: The digital microfluidic chip comprises an electrode array, the surface of the electrode array is provided with the reagent pre-embedding and sample injection device according to any one of claims 1 to 10, the surface of the electrode array is also provided with a dielectric layer and a hydrophobic layer stacked in sequence, a transparent conductive cover is buckled on the top of the hydrophobic layer, a gap cavity is formed between the transparent conductive cover and the hydrophobic layer, and the bottom liquid injection port of the sample injection seat is connected to the gap cavity.

14. The digital microfluidic chip according to claim 13, characterized in that: The reagent pre-embedding and sample injection device is fixed on the edge of the electrode array.

15. The digital microfluidic chip according to claim 13, characterized in that: At least two reagent pre-embedding and sample injection devices are fixed side by side along the same edge of the electrode array.

16. The digital microfluidic chip according to claim 13, characterized in that: The reagents stored in each reagent pack may be the same or different.

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