Three-dimensional microfluidic hazard factor detection chip

Abstract

The present invention discloses a three-dimensional microfluidic hazard factor detection chip, and relates to the technical field of microfluidics, the three-dimensional microfluidic hazard factor detection chip at least comprises a microfluidic chip main body and a detection reaction base, a magnetic rod accommodating hole, a spiral channel, a liquid adding cavity, a waste liquid cavity, a detection reaction base positioning opening, a liquid outlet, a first air suction channel and a second air suction channel are arranged in the micro-fluidic chip main body, and a detachable magnetic rod is arranged in the magnetic rod accommodating hole; the detection reaction base is detachably arranged at a detection reaction base positioning opening in the bottom of the micro-fluidic chip main body, and the detection reaction base and the detection reaction base positioning opening are in sealing fit; the invention provides a three-dimensional microfluidic hazard factor detection chip capable of integrating uniform mixing, magnetic separation, purification and one-step multi-joint detection.

Description

Technical Field

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

[0002] At present, biological hazards and chemical hazards have become the focus of global attention. The problems caused by them, such as disease health, product quality, and agricultural production, seriously affect people's life safety and cause inestimable economic losses. Different detection technologies and equipment have been developed for the detection of various hazard factors, mainly focusing on nucleic acid detection, immunoassay, electrochemical detection, and spectral detection technology systems. Based on these technologies, researchers have developed a large number of detection equipment. Such as fluorescence quantitative PCR instruments, microplate 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-machine detection, false negatives and missed detection. Especially in the face of unknown sample detection, it is often necessary to use multiple technologies and equipment in combination. How to enable multiple hazard factors to achieve rapid multi-link detection in the same detection technology system, maximize the use efficiency of samples and ensure the accuracy of test 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 achieving high-throughput, multi-target detection. However, the current mainstream planar microfluidic chips have problems such as drive control limitations, insufficient expansion space, and difficulty in modular integration, which limit 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 that can integrate mixing, magnetic separation, purification, and one-step multi-link detection.

[0005] In order to achieve the above object, the present invention adopts the following technical solutions: A three-dimensional microfluidic hazard factor detection chip comprises at least a microfluidic chip body and a detection reaction base, wherein the microfluidic chip body is columnar and is provided with: A magnetic rod receiving hole, the magnetic rod receiving hole is vertically arranged and has an open top, and a detachable magnetic rod is arranged in the magnetic rod receiving hole; A spiral channel, wherein the spiral channel is arranged around the lower part of the magnetic rod receiving hole, and the central axis of the spiral channel coincides with the central axis of the magnetic rod receiving hole; A liquid adding chamber, wherein the top of the liquid adding chamber is open, and the bottom is connected to the upper end of the spiral channel; A waste liquid chamber, the waste liquid chamber being communicated with the lower end of the spiral channel; A detection reaction base positioning port, wherein the detection reaction base positioning port is located at the bottom of the microfluidic chip body; A liquid outlet, the liquid outlet is located at the inner top of the detection reaction base positioning port, and the liquid outlet is connected to the lower end of the spiral channel; A first air suction channel, wherein one end of the first air suction channel is connected to the outer wall of the microfluidic chip body, and the other end of the first air suction channel is connected to the waste liquid chamber; A second air suction channel, one end of which is connected to the outer wall of the microfluidic chip body, and the other end of which is connected to the inner top of the positioning port of the detection reaction base; The detection reaction base is detachably arranged at the detection reaction base positioning opening at the bottom of the microfluidic chip body, and the detection reaction base is sealed and matched with the detection reaction base positioning opening.

[0006] Preferably, the spiral channel is in a conical spiral shape with a large upper opening and a small lower opening.

[0007] Preferably, the liquid outlet is a downwardly protruding nozzle structure.

[0008] 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, a sealing ring is circumferentially provided on the outer side of the cylindrical structure, the detection reaction base positioning port is cylindrical and seals with the upper part of the sealing seat, a detection array chip positioning groove is provided on the top of the sealing seat, the detection array chip is arranged in the detection array chip positioning groove, a reagent containing groove is provided on the top of the detection array chip, and the liquid outlet is located on the upper side of the reagent containing groove at the top of the detection array chip.

[0009] Preferably, the inner bottom of the detection array chip is provided with a plurality of concave single-target detection microgrooves independently distributed in an array. The detection array chip is concave, with an open top and four higher sides, and the inner side can accommodate the liquid to be detected.

[0010] Preferably, the bottom of the liquid adding chamber is funnel-shaped.

[0011] Preferably, it further comprises a waste liquid cavity plug; a waste liquid discharge port is provided on the microfluidic chip body; the waste liquid discharge port is communicated with the waste liquid cavity on the inner side and is blocked by the waste liquid cavity plug on the outer side.

[0012] Preferably, the front side of the waste liquid chamber plug is in a transverse cylindrical shape, and a sealing ring is circumferentially arranged on the outer side of the cylindrical structure, through which the waste liquid chamber plug is sealed with the waste liquid discharge port.

[0013] The present invention has the following beneficial effects: 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; 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. 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; 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; 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

[0014] Figure 1 It is a structural schematic diagram of the present invention.

[0015] Figure 2 It is a perspective view of the present invention.

[0016] Figure 3 It is an exploded view of the present invention.

[0017] Figure 4 It is an exploded view from another angle of the present invention.

[0018] Figure 5It is a perspective view of the present invention in an exploded state.

[0019] Figure 6 It is a perspective view of the exploded state of the present invention from another angle.

[0020] Figure 7 It is a top view of the main body of the microfluidic chip in the present invention.

[0021] Figure 8 yes Figure 7 Cross-sectional view along the AA direction.

[0022] 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 rod accommodating hole; 4: magnetic rod; 5: spiral channel; 6: liquid addition chamber; 7: waste liquid chamber; 8: detection reaction base positioning port; 9: liquid outlet; 10: first air suction channel; 11: second air suction channel; 12: waste liquid chamber plug; 13: waste liquid discharge port. DETAILED DESCRIPTION

[0023] The present invention will be further described below in conjunction with the accompanying drawings and specific embodiments.

[0024] A three-dimensional microfluidic hazard factor detection chip, see Figures 1 to 8 , at least comprising a microfluidic chip body 1 and a detection reaction base 2, wherein the microfluidic chip body 1 is columnar, and the microfluidic chip body 1 is provided with: A magnetic rod receiving hole 3, wherein the magnetic rod receiving hole 3 is vertically arranged and has an open top, and a detachable magnetic rod 4 is arranged in the magnetic rod receiving hole 3; A spiral channel 5, wherein the spiral channel 5 is arranged around the lower part of the magnetic rod accommodating hole 3, and the central axis of the spiral channel 5 coincides with the central axis of the magnetic rod accommodating hole 3, and the spiral channel 5 is in a conical spiral shape with a large upper opening and a small lower opening; A liquid adding chamber 6, wherein the top of the liquid adding chamber 6 is open, and the bottom of the liquid adding chamber 6 is funnel-shaped, and the bottom is connected to the upper end of the spiral channel 5; A waste liquid chamber 7, wherein the waste liquid chamber 7 is connected to the lower end of the spiral channel 5, and a connecting port between the spiral channel 5 and the waste liquid chamber 7 is located at the top of the waste liquid chamber 7; A detection reaction base positioning port 8, wherein 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 connected to the lower end of the spiral channel 5, and the liquid outlet 9 is a nozzle structure protruding downward; A first air suction channel 10, one end of which is connected to the outer wall of the microfluidic chip body 1, and the other end is connected to the waste liquid chamber 7, and the connecting port between the first air suction channel 10 and the waste liquid chamber 7 is located at the top of the waste liquid chamber 7; A second air suction channel 11, one end of which is connected to the outer wall of the microfluidic chip body 1, and the other end of which is connected 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 opening 8 at the bottom of the microfluidic chip body 1 , and the detection reaction base 2 and the detection reaction base positioning opening 8 are sealed and matched.

[0025] 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 provided in the outer side of the cylindrical structure. The detection reaction base positioning port 8 is cylindrical and sealed with the upper part of the sealing seat 201. The top of the sealing seat 201 is provided with a detection array chip positioning groove 203. The detection array chip 202 is arranged in the detection array chip positioning groove 203. The top of the detection array chip 202 is provided with a reagent containing groove. The bottom of the inner side of the reagent containing groove of the detection array chip 202 is provided with 16 concave single target detection micro grooves 204 independently distributed in a 4×4 array. The liquid outlet 9 is located on the upper side of the reagent containing groove at the top of the detection array chip 202. The detection reaction base 2 is designed to be a sealing seat 201 and a detection array chip 202, so that the sealing seat 201 will not be contaminated by contact with the reagent, and can be reused, thereby increasing the environmental protection of the product.

[0026] The present invention further includes a waste liquid cavity plug 12, and a waste liquid discharge port 13 is provided on the microfluidic chip body 1. The inside of the waste liquid discharge port 13 is connected to the waste liquid cavity 7, and the outside is blocked by the waste liquid cavity plug 12. The front side of the waste liquid cavity plug 12 is in a transverse cylindrical shape, and a sealing ring is provided in an annular direction on the outside of the cylindrical structure, and the waste liquid cavity plug 12 is sealed and matched with the waste liquid discharge port 13 through the sealing ring.

[0027] When the present invention is used to perform a hazard factor detection test, a suction device (such as a syringe) is first connected to the openings of the first suction channel 10 and the second suction channel 11, and a magnetic rod 4 is inserted into the magnetic rod receiving hole 3.

[0028] The sample solution and the modified magnetic beads (modified with nucleic acid aptamers, probes, antibodies, etc.) solution are respectively injected into the liquid adding chamber 6 to form a mixed solution. At this time, the modified magnetic beads can specifically bind to the target to be detected.

[0029] Then, the suction device on the first suction channel 10 starts the suction work, slowly sucks air, and a negative pressure is formed in the waste liquid chamber 7. The mixed liquid in the liquid adding 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 mixed liquid passes through the spiral channel 5, at first, the upper spiral channel 5 is far away from the magnetic rod 4, and the magnetism is weak, which ensures the fluidity of the mixed liquid, and the modified magnetic beads in the mixed liquid will not be concentratedly adsorbed and block the channel. At the same time, when the mixed liquid flows in the spiral channel 5, due to the combined effect of factors such as the narrow spiral channel, the long spiral distance, and the continuous change of the spiral centrifugal force, gravity and magnetic force, the contact between the target to be tested and the modified magnetic beads can be more sufficient, thereby effectively promoting the modified magnetic beads to capture the target to be tested. The mixed liquid continues to flow downward, and 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 closer to the magnetic rod 4 and has stronger magnetism, which effectively prevents the remaining unabsorbed modified magnetic beads from flowing through the spiral channel 5 and entering the waste liquid chamber 7, thereby improving the recovery rate of the magnetic beads and reducing the probability of missed detection.

[0030] When the mixed liquid in the liquid adding chamber 6 and the spiral channel 5 flows out, the modified magnetic beads in the mixed liquid are basically adsorbed on the inner wall of the spiral channel 5, and these modified magnetic beads have been fully in contact with the target to be detected, and the surfaces of many modified magnetic beads have captured the target to be detected.

[0031] Then, the channel is cleaned with an eluent to elute the substances remaining in the spiral channel 5 that are not captured by the modified magnetic beads and other impurities from the channel. A sufficient amount of eluent is added to the liquid adding chamber 6, and the suction device on the first suction channel 10 continues to suction, and the eluent in the liquid adding chamber 6 still enters the waste liquid chamber 7 through the spiral channel 5.

[0032] After the impurities are removed, the suction device on the first suction channel 10 stops working and starts to collect the modified magnetic beads: the magnetic rod 4 is pulled out from the magnetic rod accommodating hole 3, and the magnetic beads on the inner wall of the spiral channel 5 lose their magnetic fixation. A sufficient amount of eluent is added to the liquid adding chamber 6 again, and the suction device on the second suction channel 11 starts the suction work, and a negative pressure is formed in the detection reaction base positioning port 8. The eluent in the liquid adding chamber 6 is affected by the negative pressure in the detection reaction base positioning port 8, flows through the spiral channel 5 and drives the modified magnetic beads that have lost their magnetic fixation, and finally flows out from the liquid outlet 9 and drips into the detection array chip 202 below the liquid outlet 9. The target recognition reaction is completed in the detection array chip 202, and then the detection reaction base 2 is removed, and the detection array chip 202 is taken out. The unbound modified magnetic beads are washed off by washing the plate, and high-definition photography is directly performed after drying, and the detection result is determined by image recognition.

Claims

1. A three-dimensional microfluidic hazard factor detection chip, characterized in that: The microfluidic chip comprises at least a microfluidic chip body (1) and a detection reaction base (2), wherein the microfluidic chip body (1) is columnar, and the microfluidic chip body (1) is provided with: A magnetic rod receiving hole (3), the magnetic rod receiving hole (3) being arranged vertically and open at the top, and a detachable magnetic rod (4) being arranged in the magnetic rod receiving hole (3); A spiral channel (5), wherein the spiral channel (5) is arranged around the lower part of the magnetic rod accommodating hole (3), and the central axis of the spiral channel (5) coincides with the central axis of the magnetic rod accommodating hole (3); A liquid adding chamber (6), wherein the top of the liquid adding chamber (6) is open and the bottom is connected to the upper end of the spiral channel (5); A waste liquid chamber (7), wherein the waste liquid chamber (7) is connected to the lower end of the spiral channel (5); A detection reaction base positioning opening (8), wherein the detection reaction base positioning opening (8) is located at the bottom of the microfluidic chip body (1); A liquid outlet (9), the liquid outlet (9) being located at the inner top of the detection reaction base positioning opening (8), and the liquid outlet (9) being 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) being connected to the outer wall of the microfluidic chip body (1), and the other end of the first air suction channel (10) being connected to the waste liquid chamber (7); A second air intake channel (11), one end of the second air intake channel (11) being connected to the outer wall of the microfluidic chip body (1), and the other end of the second air intake channel (11) being connected to the inner top of the detection reaction base positioning opening (8); The detection reaction base (2) is detachably arranged at a detection reaction base positioning opening (8) at the bottom of the microfluidic chip body (1), and the detection reaction base (2) and the detection reaction base positioning opening (8) are sealed and matched.

2. A three-dimensional microfluidic hazard factor detection chip according to claim 1, characterized in that: The spiral channel (5) is in the shape of a cone spiral with a large upper opening and a small lower opening.

3. A three-dimensional microfluidic hazard factor detection chip according to claim 1, characterized in that: The liquid outlet (9) is a nozzle structure protruding downward.

4. A three-dimensional microfluidic hazard factor detection chip according to claim 1 or 3, characterized in that: The detection reaction base (2) is composed of a sealing seat (201) and a detection array chip (202); the upper portion of the sealing seat (201) is cylindrical, a sealing ring is provided in an annular direction on the outer side of the cylindrical structure, the detection reaction base positioning opening (8) is cylindrical and seals with the upper portion of the sealing seat (201); the top of the sealing seat (201) is provided with a detection array chip positioning groove (203), the detection array chip (202) is arranged in the detection array chip positioning groove (203), the top of the detection array chip (202) is provided with a reagent containing groove, and the liquid outlet (9) is located on the upper side of the reagent containing groove at the top of the detection array chip (202).

5. A three-dimensional microfluidic hazard factor detection chip according to claim 4, characterized in that: The inner bottom of the detection array chip (202) is provided with a plurality of concave single-target detection microgrooves (204) which are independently distributed in an array.

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

7. The three-dimensional microfluidic hazard factor detection chip according to claim 1, characterized in that: It also includes a waste liquid cavity plug (12), and a waste liquid discharge port (13) is provided on the microfluidic chip body (1). The inside of the waste liquid discharge port (13) is connected to the waste liquid cavity (7), and the outside is blocked by the waste liquid cavity plug (12).

8. A three-dimensional microfluidic hazard factor detection chip according to claim 7, characterized in that: The front side of the waste liquid cavity plug (12) is in the shape of a horizontal cylinder, and a sealing ring is provided in an annular direction on the outer side of the cylindrical structure, through which the waste liquid cavity plug (12) is sealed and matched with the waste liquid discharge port (13).

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