A three-dimensional microfluidic hazard factor detection chip

Abstract

The present invention discloses a three-dimensional microfluidic hazard factor detection chip, which relates to the field of microfluidic technology and at least includes a microfluidic chip main body and a detection reaction base. The microfluidic chip main body is columnar, and the microfluidic chip main body is provided with: a magnetic bar accommodation hole, a spiral channel, a liquid addition chamber, a waste liquid chamber, a detection reaction base positioning port, a liquid outlet, a first suction channel and a second suction channel. A detachable magnetic bar is arranged in the magnetic bar accommodation hole; the detection reaction base is detachably arranged at the detection reaction base positioning port at the bottom of the microfluidic chip main body, and a sealed fit is provided between the detection reaction base and the detection reaction base positioning port; the present invention provides a three-dimensional microfluidic hazard factor detection chip that can integrate mixing, magnetic separation, purification, and one-step multiplex detection.

Description

Technical Field

[0001] The present invention relates to the field of microfluidic technology, and particularly to a hazard factor detection chip combined with three-dimensional microfluidics. Background Art

[0002] For the detection of various hazard factors, different detection technologies and devices have been developed, mainly concentrated in the nucleic acid detection, immunoassay, electrochemical detection, and spectroscopic detection technology systems. Based on these technologies, researchers have developed a large number of detection devices. Such as fluorescence quantitative PCR instruments, enzyme-linked immunosorbent assay (ELISA) readers, electrochemical workstations, Raman detectors, etc. The application of these instruments has played a positive role in the research and prevention and control of hazard factors, but there are still problems such as cumbersome operation, expensive equipment, multi-instrument detection, false negative and missed detection. Especially when facing the detection of unknown samples, multiple technologies and devices are often required to be used in combination. How to enable multiple hazard factors to achieve rapid multiplex detection in the same detection technology system, maximize the use efficiency of samples and ensure the accuracy of detection results is an urgent problem to be solved.

[0003] The detection method based on microfluidic technology has the characteristics of anti-pollution, integration, low cost, fast analysis speed, and parallel detection, and is an ideal strategy for realizing high-throughput and multi-target detection. However, the current mainstream planar microfluidic chips have problems such as driving control limitations, insufficient expandable space, and difficulty in modular integration, resulting in limitations in their use functions and scenarios. Summary of the Invention

[0004] In order to solve the above problems, the present invention proposes a three-dimensional microfluidic hazard factor detection chip capable of integrating mixing, magnetic separation, purification, and one-step multiplex detection.

[0005] In order to achieve the above object, the present invention adopts the following technical solutions:

[0006] A three-dimensional microfluidic hazard factor detection chip, at least including a microfluidic chip main body and a detection reaction base, the microfluidic chip main body is columnar, and the microfluidic chip main body is provided with:

[0007] A magnetic rod accommodating hole, the magnetic rod accommodating hole is vertically arranged and has an open top, and a detachable magnetic rod is arranged in the magnetic rod accommodating hole;

[0008] A spiral channel, the spiral channel is arranged around the lower part of the magnetic rod accommodating hole, and the central axis of the spiral channel coincides with the central axis of the magnetic rod accommodating hole;

[0009] A liquid adding cavity, the liquid adding cavity has an open top and its bottom is communicated with the upper end of the spiral channel;

[0010] A waste liquid cavity, the waste liquid cavity is communicated with the lower end of the spiral channel;

[0011] Detect the positioning port of the reaction base, and the positioning port of the reaction base is located at the bottom of the microfluidic chip body;

[0012] The liquid outlet is located at the inner top of the positioning port of the detection reaction base, and the liquid outlet is communicated with the lower end of the spiral channel;

[0013] The first suction channel, one end of the first suction channel is communicated with the outer wall of the microfluidic chip body, and the other end is communicated with the waste liquid chamber;

[0014] The second suction channel, one end of the second suction channel is communicated with the outer wall of the microfluidic chip body, and the other end is communicated with the inner top of the positioning port of the detection reaction base;

[0015] The detection reaction base is detachably arranged at the positioning port of the detection reaction base at the bottom of the microfluidic chip body, and the detection reaction base and the positioning port of the detection reaction base are in sealing fit.

[0016] Preferably, the spiral channel is in a conical spiral shape with a large upper opening and a small lower opening.

[0017] Preferably, the liquid outlet is a nozzle structure protruding downward.

[0018] Preferably, the detection reaction base is composed of a sealing seat and a detection array chip. The upper part of the sealing seat is cylindrical, and a sealing ring is circumferentially arranged on the outer side of the cylindrical structure. The positioning port of the detection reaction base is cylindrical and is in sealing fit with the upper part of the sealing seat. A detection array chip positioning groove is arranged on the top of the sealing seat. The detection array chip is arranged in the detection array chip positioning groove. A reagent accommodating groove is arranged on the top of the detection array chip, and the liquid outlet is located above the reagent accommodating groove on the top of the detection array chip.

[0019] Preferably, a plurality of concave single-target detection micro-grooves are arranged in an array and independently distributed at the inner bottom of the detection array chip. The detection array chip is concave, with an open top, relatively high sides, and can accommodate the liquid to be tested inside.

[0020] Preferably, the bottom of the liquid adding chamber is in a funnel shape.

[0021] Preferably, it further includes a waste liquid chamber plug. A waste liquid discharge port is arranged on the microfluidic chip body. The inner side of the waste liquid discharge port is communicated with the waste liquid chamber, and the outer side is blocked by the waste liquid chamber plug.

[0022] Preferably, the front side of the waste liquid chamber plug is in a horizontal cylindrical shape, and a sealing ring is circumferentially arranged on the outer side of the cylindrical structure. The waste liquid chamber plug is in sealing fit with the waste liquid discharge port through the sealing ring.

[0023] The present invention has the following beneficial effects:

[0024] 1. A spiral channel is provided inside, so that when the solution containing the sample to be tested and the mixed solution containing the modified magnetic beads such as probes / antibodies move downward in the spiral channel, under the continuous changes of the spiral centrifugal force, gravity and magnetic force, the modified magnetic beads and the target to be tested can be fully mixed and contacted in the longer spiral channel, effectively improving the capture rate of the magnetic beads;

[0025] 2. The spiral channel is set to a conical spiral shape with a large upper opening and a small lower opening. The upper spiral channel is far away from the magnetic rod and has weaker magnetism, which ensures the fluidity of the mixed solution and prevents the magnetic beads from being concentratedly adsorbed and blocking the channel. As the distance to the magnetic rod approaches, the magnetism becomes stronger and the magnetic beads are gradually adsorbed on the inner wall of the spiral channel. The lower spiral channel is close to the magnetic rod and has stronger magnetism, which prevents the remaining unadsorbed magnetic beads from flowing through the spiral channel and entering the waste liquid chamber, causing missed detection of the target to be tested.

[0026] 3. The magnetic rod adopts a pluggable design. After taking out the magnetic rod, the magnetic beads adsorbed on the inner wall of the spiral channel can be eluted by the eluent and dripped into the detection array chip on the detection reaction base for specific recognition reaction. The test liquid soaks all the concave single-target detection microgrooves, and the target to be tested combined with the modified magnetic beads specifically binds to the probe / antigen in the concave single-target detection microgrooves with the same probability. After incubation at constant temperature with shaking for a period of time, take out the detection array chip and wash the plate to wash away the unbound magnetic beads. After drying, high-definition photography and imaging are directly performed, and the detection results are determined by image recognition;

[0027] 4. Each concave single-target detection microgroove can be coated with different detection probes / antibodies according to the detection needs, so as to achieve the simultaneous detection of different types of harmful factors, and realize accurate identification through spatial encoding through the detection array chip, without the need for complex operations such as multiple fluorescent labels;

[0028] 5. The microfluidic chip will drop the sample after efficient magnetic separation and enrichment and purification directly into the detection array chip and cover all concave single-target detection microgrooves. All target magnetic bead complexes to be tested will contact the coated probe / antibody in the detection microgrooves with the same probability. The function of one-time multi-target detection can be realized without liquid separation and diversion of samples, reducing missed detection and false negatives. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 It is a structural schematic diagram of the present invention.

[0030] Figure 2 It is a perspective view of the present invention.

[0031] Figure 3 It is an exploded view of the present invention.

[0032] Figure 4It is an exploded view from another angle of the present invention.

[0033] Figure 5 It is a perspective view of the present invention in an exploded state.

[0034] Figure 6 It is a perspective view of the present invention in an exploded state from another angle.

[0035] Figure 7 It is a top view of the microfluidic chip body in the present invention.

[0036] Figure 8 It is Figure 7 a cross-sectional view taken along the A-A direction in

[0037] 1: Microfluidic chip body; 2: Detection reaction base; 201: Sealing seat; 202: Detection array chip; 203: Detection array chip positioning groove; 204: Concave single-target detection microgroove; 3: Magnetic bar accommodating hole; 4: Magnetic bar; 5: Spiral channel; 6: Liquid addition chamber; 7: Waste liquid chamber; 8: Detection reaction base positioning port; 9: Liquid outlet; 10: First suction channel; 11: Second suction channel; 12: Waste liquid chamber plug; 13: Waste liquid discharge port. Detailed implementation manners

[0038] The following further describes the present invention in conjunction with the drawings and specific implementation manners.

[0039] A three-dimensional microfluidic hazard factor detection chip, referring to Figures 1 to 8 , at least includes a microfluidic chip body 1 and a detection reaction base 2. The microfluidic chip body 1 is columnar, and inside the microfluidic chip body 1 are provided with:

[0040] A magnetic bar accommodating hole 3, the magnetic bar accommodating hole 3 is vertically arranged and has an open top, and a detachable magnetic bar 4 is arranged inside the magnetic bar accommodating hole 3;

[0041] A spiral channel 5, the spiral channel 5 is arranged around the lower part of the magnetic bar accommodating hole 3, and the central axis of the spiral channel 5 coincides with the central axis of the magnetic bar accommodating hole 3. The spiral channel 5 is in a conical spiral shape with a large upper opening and a small lower opening;

[0042] A liquid addition chamber 6, the liquid addition chamber 6 has an open top, and the bottom of the liquid addition chamber 6 is funnel-shaped and communicates with the upper end of the spiral channel 5 at the bottom;

[0043] A waste liquid chamber 7, the waste liquid chamber 7 communicates with the lower end of the spiral channel 5, and the communication port of the spiral channel 5 and the waste liquid chamber 7 is located at the top of the waste liquid chamber 7;

[0044] A detection reaction base positioning port 8, the detection reaction base positioning port 8 is located at the bottom of the microfluidic chip body 1;

[0045] A liquid outlet 9, the liquid outlet 9 is located at the inner top of the detection reaction base positioning port 8, and the liquid outlet 9 communicates with the lower end of the spiral channel 5. The liquid outlet 9 is a nozzle structure protruding downward;

[0046] A first suction channel 10, one end of the first suction channel 10 communicates with the outer wall of the microfluidic chip body 1, the other end communicates with the waste liquid chamber 7, and the communication port of the first suction channel 10 and the waste liquid chamber 7 is located at the top of the waste liquid chamber 7;

[0047] A second suction channel 11, one end of the second suction channel 11 communicates with the outer wall of the microfluidic chip body 1, and the other end communicates with the inner top of the detection reaction base positioning port 8;

[0048] The detection reaction base 2 is detachably arranged at the detection reaction base positioning port 8 at the bottom of the microfluidic chip body 1, and the detection reaction base 2 and the detection reaction base positioning port 8 are in sealing fit.

[0049] The detection reaction base 2 is composed of a sealing seat 201 and a detection array chip 202. The upper part of the sealing seat 201 is cylindrical, and a sealing ring is arranged circumferentially on the outer side of the cylindrical structure. The detection reaction base positioning port 8 is cylindrical and is in sealing fit with the upper part of the sealing seat 201. A detection array chip positioning groove 203 is arranged at the top of the sealing seat 201. The detection array chip 202 is arranged in the detection array chip positioning groove 203. A reagent accommodating groove is arranged at the top of the detection array chip 202. Sixteen concave single-target detection micro-grooves 204 distributed independently in a 4×4 array are arranged at the inner bottom of the reagent accommodating groove of the detection array chip 202. The liquid outlet 9 is located above the reagent accommodating groove at the top of the detection array chip 202. Designing the detection reaction base 2 into two parts, namely the sealing seat 201 and the detection array chip 202, enables the sealing seat 201 not to be contaminated by the reagent and can be reused, increasing the environmental protection of the product.

[0050] The present invention further includes a waste liquid chamber plug 12. A waste liquid discharge port 13 is arranged on the microfluidic chip body 1. The inner side of the waste liquid discharge port 13 communicates with the waste liquid chamber 7, and the outer side is blocked by the waste liquid chamber plug 12. The front side of the waste liquid chamber plug 12 is horizontally cylindrical, and a sealing ring is arranged circumferentially on the outer side of the cylindrical structure. The waste liquid chamber plug 12 is in sealing fit with the waste liquid discharge port 13 through the sealing ring.

[0051] When using the present invention to conduct a hazard factor detection test, first connect a suction device (such as a syringe) to the openings of the first suction channel 10 and the second suction channel 11, and insert a magnetic bar 4 into the magnetic bar accommodating hole 3.

[0052] The sample solution and the solution of modified magnetic beads (modified with nucleic acid aptamers, probes, antibodies, etc.) are respectively injected into the liquid addition chamber 6 to form a mixture. At this time, the modified magnetic beads can specifically bind to the target to be detected.

[0053] Then, the suction device on the first suction channel 10 starts to work for suction, sucking slowly, a negative pressure is formed in the waste liquid chamber 7, and the mixture in the liquid addition chamber 6 is affected by the negative pressure in the waste liquid chamber 7 and will slowly pass through the spiral channel 5 and enter the waste liquid chamber 7. When the mixture passes through the spiral channel 5, at first, the upper spiral channel 5 is far from the magnetic rod 4 and has a weak magnetism, ensuring the fluidity of the mixture. The modified magnetic beads in the mixture will not be concentrated and adsorbed to block the channel. At the same time, when the mixture flows in the spiral channel 5, due to the combined action of factors such as the narrow spiral channel, long spiral distance, and continuous change of spiral centrifugal force, gravity, and magnetism, the contact between the target to be detected and the modified magnetic beads can be made more sufficient, thereby effectively promoting the capture of the target to be detected by the modified magnetic beads. The mixture continues to flow downward. As the distance between the spiral channel 5 and the magnetic rod approaches, the magnetism becomes stronger, and the modified magnetic beads are gradually adsorbed on the inner wall of the spiral channel 5. The lower spiral channel 5 is close to the magnetic rod 4 and has a strong magnetism, effectively preventing the remaining unadsorbed modified magnetic beads from flowing through the spiral channel 5 and entering the waste liquid chamber 7, improving the recovery rate of the magnetic beads and reducing the probability of missed detection.

[0054] When the mixture in the liquid addition chamber 6 and the spiral channel 5 has flowed out, the modified magnetic beads in the mixture are basically adsorbed on the inner wall of the spiral channel 5, and these modified magnetic beads have made sufficient contact with the target to be detected, and many modified magnetic beads have captured the target to be detected on their surfaces.

[0055] Then, the channel is washed with the eluent to elute the substances that have not been captured by the modified magnetic beads and other impurities remaining in the spiral channel 5 from the channel. A sufficient amount of eluent is added to the liquid addition chamber 6, and the suction device on the first suction channel 10 continues to work for suction, and the eluent in the liquid addition chamber 6 still enters the waste liquid chamber 7 through the spiral channel 5.

[0056] After the impurities are removed, the suction device on the first suction channel 10 stops working, and the collection of modified magnetic beads begins: the magnetic rod 4 is withdrawn from the magnetic rod accommodating hole 3, and the magnetic beads on the inner wall of the spiral channel 5 lose magnetic fixation. Again, a sufficient amount of eluent is added into the liquid adding chamber 6, and the suction device on the second suction channel 11 starts suction work. It is detected that a negative pressure is formed in the positioning port 8 of the reaction base. The eluent in the liquid adding chamber 6 is affected by the negative pressure in the positioning port 8 of the reaction base, flows through the spiral channel 5 and drives the modified magnetic beads that have lost magnetic fixation to flow together, and finally flows out from the liquid outlet 9 and drops into the detection array chip 202 below the liquid outlet 9. The target recognition reaction is completed in the detection array chip 202, then the detection reaction base 2 is removed, the detection array chip 202 is taken out, the unbound modified magnetic beads are washed away through plate washing, and after drying, high-definition photography imaging is directly carried out, and the detection result is judged through image recognition.

Claims

1. A three-dimensional microfluidic hazard factor detection chip, characterized in that, It includes at least a microfluidic chip body (1) and a detection reaction base (2). The microfluidic chip body (1) is columnar, and inside the microfluidic chip body (1) are provided with: A magnetic bar accommodating hole (3), the magnetic bar accommodating hole (3) is vertically arranged and has an open top, and a detachable magnetic bar (4) is provided inside the magnetic bar accommodating hole (3); A spiral channel (5), the spiral channel (5) is arranged around the lower part of the magnetic bar accommodating hole (3), and the central axis of the spiral channel (5) coincides with the central axis of the magnetic bar accommodating hole (3); the spiral channel (5) is in a conical spiral shape with a larger opening at the upper part and a smaller opening at the lower part; A liquid adding cavity (6), the liquid adding cavity (6) has an open top and its bottom is communicated with the upper end of the spiral channel (5); A waste liquid cavity (7), the waste liquid cavity (7) is communicated with the lower end of the spiral channel (5); A detection reaction base positioning port (8), the detection reaction base positioning port (8) is located at the bottom of the microfluidic chip body (1); A liquid outlet (9), the liquid outlet (9) is located at the inner top of the detection reaction base positioning port (8), and the liquid outlet (9) is communicated with the lower end of the spiral channel (5); A first air suction channel (10), one end of the first air suction channel (10) is communicated to the outer wall of the microfluidic chip body (1), and the other end is communicated with the waste liquid cavity (7), and the communication port of the first air suction channel (10) and the waste liquid cavity (7) is located at the top of the waste liquid cavity (7); A second air suction channel (11), one end of the second air suction channel (11) is communicated to the outer wall of the microfluidic chip body (1), and the other end is communicated to the inner top of the detection reaction base positioning port (8); The detection reaction base (2) is detachably arranged at the detection reaction base positioning port (8) at the bottom of the microfluidic chip body (1), and there is a sealing fit between the detection reaction base (2) and the detection reaction base positioning port (8); the detection reaction base (2) is composed of a sealing seat (201) and a detection array chip (202). The upper part of the sealing seat (201) is cylindrical, and an O-ring is circumferentially arranged on the outer side of this cylindrical structure. The detection reaction base positioning port (8) is cylindrical and is in sealing fit with the upper part of the sealing seat (201). A detection array chip positioning groove (203) is provided at the top of the sealing seat (201), the detection array chip (202) is arranged in the detection array chip positioning groove (203), a reagent accommodating groove is provided at the top of the detection array chip (202), and the liquid outlet (9) is located above the reagent accommodating groove at the top of the detection array chip (202).

2. The three-dimensional microfluidic hazard factor detection chip according to claim 1, wherein: The liquid outlet (9) is a nozzle structure protruding downward.

3. The three-dimensional microfluidic hazard factor detection chip according to claim 1, wherein: Several concave single-target detection micro-grooves (204) distributed independently in an array are provided at the inner bottom of the detection array chip (202).

4. A three-dimensional microfluidic hazard factor detection chip according to claim 1, characterized in that: The bottom of the liquid adding cavity (6) is in a funnel shape.

5. A three-dimensional microfluidic hazard factor detection chip according to claim 1, characterized in that: It further includes a waste liquid cavity plug (12). A waste liquid discharge port (13) is provided on the microfluidic chip body (1). The inner side of the waste liquid discharge port (13) is communicated with the waste liquid cavity (7), and the outer side is blocked by the waste liquid cavity plug (12).

6. The three-dimensional microfluidic hazard factor detection chip according to claim 5, characterized in that: The front side of the waste liquid chamber plug (12) is in a horizontal cylindrical shape, and a sealing ring is circumferentially arranged on the outer side of the cylindrical structure. The waste liquid chamber plug (12) is hermetically fitted with the waste liquid discharge port (13) through the sealing ring.

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