Biomolecule multiple detection chip, detection assembly and detection system based on micro flow field and micro magnetic field coupling

Abstract

The utility model discloses a biological molecule multiple detection chip, detection subassembly and detection system based on micro -flow field and micro -magnetic field coupling action, and the chip includes chip substrate, and the top of chip substrate is provided with first liquid inlet, second liquid inlet, first liquid outlet and second liquid outlet, and the inside of chip substrate is provided with ball trap flow channel, detection flow channel and capture cavity, and first liquid inlet is used for corresponding and is passed into different type's functionalized magnetic microsphere solution, and second liquid inlet is used for passing in the sample solution of detection and detection solution, and every capture cavity is communicated in proper order through detection flow channel, and the micro -magnetic cone is embedded in the chip substrate below capture cavity, and the detection subassembly includes: based on chip and setting below chip, and the magnet of adjustable spacing with micro -magnetic cone, and the detection system includes: the liquid accumulator of liquid inlet intercommunication, and the waste liquid pool of import and liquid outlet intercommunication, and the negative pressure pump of waste liquid pool's outlet intercommunication and the light source for irradiating magnetic microsphere.

Description

Technical Field

[0001] This invention belongs to the field of biochip detection technology, and particularly relates to a biomolecule multiplex detection chip, detection component and detection system based on the coupling effect of microfluidic field and micromagnetic field. Background Technology

[0002] The detection of biomolecules is of great significance for research and clinical applications in the biological field. The results of biomolecule detection can serve as important clinical indicators to guide patient treatment planning. By detecting the levels of biomolecules such as nucleic acids and proteins in the human body, it is possible to diagnose related diseases, thus helping patients to detect and treat these diseases as early as possible. For example, when the level of alpha-fetoprotein (AFP) in the human body is greater than 200 ng / ml, the patient is highly likely to have primary liver cancer. In addition, the detection of biomolecules can also serve as an important indicator for postoperative monitoring of patients.

[0003] With the continuous development of biomolecular detection technology, people are paying increasing attention to the multiplexing of biomolecules. However, most current high-sensitivity and high-specificity biomolecular detection methods can only detect one type of biomolecule, and they still have certain limitations when detecting multiple biomolecules simultaneously, such as enzyme-linked immunosorbent assay (ELISA) and immunofluorescence technique (IF). Although some multiplexing methods for biomolecules exist, such as mass spectrometry (MS) and flow cytometry-encoded microsphere array (CBA) technology, they require relatively expensive instruments and have complex operating procedures, making them unsuitable for most application scenarios. Therefore, it is very important to develop a simple, low-cost, and rapid multiplexing chip for biomolecules. Summary of the Invention

[0004] To address the aforementioned issues, it is necessary to provide a biomolecular multiplex detection chip, detection component, and detection system based on the coupling effect of microfluidic field and micromagnetic field.

[0005] The first aspect of this utility model provides a biomolecular multiplex detection chip based on the coupling effect of microfluidic field and micromagnetic field, including a chip substrate;

[0006] The top of the chip substrate is provided with a first liquid inlet, a second liquid inlet, a first liquid outlet, and a second liquid outlet;

[0007] The chip substrate has a ball-catching channel, a detection channel, and multiple capture cavities inside.

[0008] The first liquid inlet is configured as a plurality of the same number as the capture chamber, for correspondingly introducing different types of functionalized magnetic microsphere solutions; the second liquid inlet is used to introduce the sample solution to be tested and the detection solution.

[0009] The first inlet of each capture chamber is connected to a first liquid inlet through the ball-catching channel, and the first outlet is connected to the first liquid outlet through the ball-catching channel.

[0010] Each capture chamber is connected sequentially through the detection channel, wherein the second inlet of the first capture chamber is connected to the second liquid inlet through the detection channel, and the second outlet of the last capture chamber is connected to the second liquid outlet.

[0011] The width of the ball-catching channel and the capturing cavity is greater than the diameter of the magnetic microspheres, and the width of the detection channel is less than the diameter of the magnetic microspheres;

[0012] A micro magnetic cone is embedded in the chip substrate below the capture cavity to capture magnetic microspheres within the capture cavity.

[0013] Based on the above, the micro magnetic cone is a magnetic nanoparticle aggregated into a cone shape.

[0014] Based on the above, the width of the ball-catching channel is greater than the diameter of the magnetic microsphere, and the width of the detection channel is less than the diameter of the magnetic microsphere.

[0015] Based on the above, the chip substrate includes an upper substrate and a lower substrate bonded together.

[0016] The ball-catching channel, the detection channel, and the capture chamber are formed on the upper substrate, and the bottom wall is open.

[0017] The micro magnetic cone is disposed on the lower substrate.

[0018] A second aspect of this invention provides a biomolecular multiplex detection component based on the coupling effect of microfluidic field and micromagnetic field, comprising:

[0019] As described in the first aspect, a biomolecular multiplex detection chip based on the coupling effect of microfluidic field and micromagnetic field; and

[0020] A magnet is positioned below the biomolecular multiplex detection chip based on the coupling effect of microfluidic field and micromagnetic field, and the distance between the magnet and the micromagnetic cone can be adjusted.

[0021] Based on the above, the magnet is a permanent magnet or an electromagnet.

[0022] Based on the above, the width range of the ball-catching channel is (3d, 4d), where d represents the diameter of the magnetic microspheres.

[0023] The third aspect of this utility model provides a biomolecular multiple detection system based on the coupling effect of microflow field and micromagnetic field, including a first liquid reservoir that is respectively connected to each first liquid inlet and is used to store different types of functionalized magnetic microsphere solutions.

[0024] A second reservoir connected to the second inlet and used for the sample solution to be tested and the detection solution;

[0025] The inlet is connected to the first outlet and is used to store waste liquid in a first waste liquid tank.

[0026] A first negative pressure pump connected to the outlet of the first waste liquid tank;

[0027] The inlet is connected to the second outlet and is used to store waste liquid in a second waste liquid tank.

[0028] A second negative pressure pump connected to the outlet of the second waste liquid tank; and

[0029] A light source used to irradiate magnetic microspheres so that the presence of fluorescent magnetic microspheres can be determined by fluorescence signal detection.

[0030] Based on the above, the bottom of the chip substrate is provided with an elastic valve for opening and closing the first liquid inlet and the first liquid outlet below the first liquid inlet and the first liquid outlet.

[0031] This utility model has substantial features and progress compared to the prior art, specifically:

[0032] 1. This utility model is equipped with multiple independent ball-catching channels. Different types of functionalized magnetic microsphere solutions enter the corresponding capture chamber from the corresponding first inlet through the corresponding ball-catching channel. The corresponding type of magnetic microspheres are captured in the corresponding capture chamber. After detection by a single fluorescence signal detection method, it can be determined whether the corresponding target analyte is present in the captured magnetic microspheres. That is, multiple detection of biomolecules can be completed in a single experiment.

[0033] 2. This utility model is provided with a ball-catching channel, a capture cavity, a detection channel and a micro magnetic cone. By utilizing the coupling effect between the microflow field formed by the ball-catching channel, the capture cavity and the detection channel and the micro magnetic field provided by the micro magnetic cone, the automatic capture and release of individual magnetic microspheres in different capture cavities is realized. Attached Figure Description

[0034] Figure 1 This is an assembly structure diagram of the biomolecular multiplex detection chip based on the coupling effect of microfluidic field and micromagnetic field in this utility model.

[0035] Figure 2 This is an exploded structural diagram of the biomolecular multiplex detection chip based on the coupling effect of microfluidic field and micromagnetic field in this utility model.

[0036] Figure 3 This invention includes a cross-sectional view of the upper and lower substrates at the capture cavity, and a schematic diagram of the magnetic field between the magnet and the micro-magnetic cone.

[0037] Figure 4 This is a detailed implementation diagram of the detection system in this utility model. Detailed Implementation

[0038] To better understand the above-mentioned objectives, features, and advantages of this utility model, the present invention will be further described in detail below with reference to the accompanying drawings and specific embodiments. It should be noted that, unless otherwise specified, the embodiments and features described in these embodiments can be combined with each other.

[0039] Many specific details are set forth in the following description in order to provide a full understanding of the present invention. However, the present invention may also be implemented in other ways different from those described herein. Therefore, the scope of protection of the present invention is not limited to the specific embodiments disclosed below.

[0040] Example 1

[0041] like Figure 1 , Figure 2 , Figure 3 , Figure 4 As shown in the figure, this embodiment proposes a biomolecular multiplex detection chip based on the coupling effect of microfluidic field and micromagnetic field.

[0042] The biomolecule multiplex detection chip includes a chip substrate 1. The top of the chip substrate 1 is provided with a first liquid inlet 102, a second liquid inlet 101, a first liquid outlet 20 and a second liquid outlet 13. The chip substrate 1 is provided with a ball-catching channel 51, a detection channel 52 and three capture chambers 53 inside.

[0043] Each capture chamber has four openings: top, bottom, left, and right. The top opening is called the first inlet, the bottom opening is called the first outlet, the right opening is called the second inlet, and the left opening is called the second outlet.

[0044] The first liquid inlet 102 is configured with three liquid inlets, namely liquid inlet 17, liquid inlet 18 and liquid inlet 19, for correspondingly introducing different types of functionalized magnetic microsphere solutions;

[0045] The second inlet 101 is used to introduce the sample solution to be tested and the detection solution. It should be noted that the second inlet 101 can be a single inlet or multiple inlets. When set as a single inlet, the sample solution to be tested and the detection solution can be introduced in stages according to the sequence of the detection steps. When set as multiple inlets, the sample solution to be tested and the detection solution can be introduced from different inlets according to the sequence of the detection steps. In this embodiment, the second inlet 101 is set to three, namely inlet 14, inlet 15, and inlet 16.

[0046] The first outlet 20 is used to discharge the waste liquid of the functionalized magnetic microsphere solution. It should be noted that the first outlet 20 can be a single outlet or multiple outlets. When multiple outlets are used, waste liquids of different types of functionalized magnetic microsphere solutions are discharged from the multiple outlets respectively. When a single outlet is used, waste liquids of different types of functionalized magnetic microsphere solutions are collected and discharged from the single outlet. In this embodiment, the first outlet 20 is set to a single outlet.

[0047] The second outlet 13 is configured as a single outlet for outputting the sample solution to be tested and the test solution.

[0048] The first inlet of the first capture chamber 53 is connected to the liquid inlet 17 through the ball-catching channel 51, the first inlet of the second capture chamber 53 is connected to the liquid inlet 18 through the ball-catching channel 51, the first inlet of the third capture chamber 53 is connected to the liquid inlet 19 through the ball-catching channel 51, and the first outlet of the three capture chambers 53 is connected to the first liquid outlet 20 through the ball-catching channel 51.

[0049] The second inlet 101 is connected to the second inlet of the third capture chamber through the detection channel 51. The second outlet of the third capture chamber 53 is connected to the second inlet of the second capture chamber 53 through the detection channel 51. The second outlet of the second capture chamber 53 is connected to the second inlet of the first capture chamber 53 through the detection channel 51. The second outlet of the first capture chamber 53 is connected to the second outlet 13.

[0050] The widths of the ball-catching channel 52 and the capturing cavity 53 are greater than the diameter of the magnetic microspheres, while the width of the detection channel 51 is smaller than the diameter of the magnetic microspheres. Micro-magnetic cones 6, 7, and 8 are embedded below the three capturing cavities 53 within the chip substrate 1 to capture the magnetic microspheres within the capturing cavities 53. In this embodiment, the magnetic microspheres cannot flow in the detection channel 51; therefore, they are only captured by the micro-magnetic cones in their respective ball-catching channels 52, thus enabling different micro-magnetic cones to capture different types of magnetic microspheres.

[0051] Specifically, the micro-magnetic cone is a cluster of magnetic nanoparticles arranged in a cone shape. Each micro-magnetic cone forms a directional connection with a magnetic microsphere through the action of a magnetic field. The tip of the cone can point towards or away from the trapping cavity 53. It should be noted that the magnetic nanoparticles are preferably magnetite nanoparticles; however, in other embodiments, magnetic nanoparticles of other materials may also be used.

[0052] Furthermore, the chip substrate 1 includes an upper substrate 3 and a lower substrate 4 bonded together; the ball-catching channel 52, the detection channel 51 and the capture cavity 53 are formed on the upper substrate 3, and the bottom wall is open; the micro magnetic cones 6, 7 and 8 are disposed on the lower substrate 4.

[0053] Figure 3 It shows a cross-sectional view of the upper substrate 2 and the lower substrate 3 at the capture cavity, as well as a schematic diagram of the magnetic field between the magnet and the micro magnetic cone. In the upper substrate 3, a detection channel 52 (transverse channel) and three ball-capturing channels 51 (longitudinal channels) intersect, and the intersection forms a capture cavity, which together form a microflow field 5. The micro magnetic cones 6, 7 and 8 in the microflow field 5 each form a magnetic field, and form directional connections with the magnetic microspheres 31, 32 and 33 through the magnetic field to complete the capture.

[0054] The working principle of the micro magnetic cone in this embodiment is as follows:

[0055] The magnetic material of a microcone is divided into many tiny regions, each called a magnetic domain, and each domain has its own magnetic moment. Before magnetization, the magnetic moments of the domains are in different directions, and the magnetic fields cancel each other out, so the entire material does not exhibit magnetism. The stronger the external magnetic field, the more these magnetic moments tend to align, and the stronger the magnetic field generated by magnetization, and vice versa.

[0056] As the strength of the external magnetic field weakens from near to far, increasing (decreasing) the distance between the magnet and the micro-magnetic cone reduces (increases) the external field strength, thereby weakening (strengthening) the tendency for the magnetic moments within the micro-magnetic cone to align, thus weakening (strengthening) the localized micro-magnetic field strength. Therefore, controlling the distance between the magnet and the micro-magnetic cone can adjust the magnetic field strength of the localized micro-magnetic field. Placing the magnet at the theoretical distance from the micro-magnetic cone generates a localized micro-magnetic field with the theoretical magnetic field strength. After removing the magnet, the micro-magnetic cone is no longer magnetized by the external magnetic field, the magnetic moments of the internal magnetic domains regain their anisotropy and cancel each other out, the localized micro-magnetic field disappears, and the magnetic microspheres are released.

[0057] Example 2

[0058] This embodiment provides a biomolecular multiplex detection component based on the coupling effect of microfluidic field and micromagnetic field, including:

[0059] The biomolecule multiplex detection chip based on the coupling effect of microfluidic field and micromagnetic field as described in Example 1; and the magnet 2 disposed below the biomolecule multiplex detection chip based on the coupling effect of microfluidic field and micromagnetic field, and whose spacing with the micromagnetic cone 6, micromagnetic cone 7, and micromagnetic cone 8 can be adjusted.

[0060] Specifically, magnet 2 is a permanent magnet or an electromagnet.

[0061] In this embodiment, magnet 2 is placed below the capture cavity 53. An external magnetic field is applied to magnet 2, and each micro-magnetic cone generates a micro-magnetic field perpendicular to the capture cavity 53 and pointing upwards under the induction of the external magnetic field. Micro-magnetic cones 6, 7, and 8 generate magnetism and can capture magnetic microspheres. During use, magnetic microspheres entering the capture cavity 53 are captured by the corresponding micro-magnetic cone. After detection, the magnet is removed, the localized micro-magnetic field disappears, and the magnetic microspheres lose their ability to capture them, releasing the magnetic microspheres.

[0062] Specifically, for ease of subsequent detection, the micro-magnetic cone is optimally suited for capturing individual magnetic microspheres. To achieve in-situ capture of a single magnetic microsphere, the base diameter of the micro-magnetic cone (taking 50 μm as an example) is related to the diameter of the magnetic microsphere (taking 50 μm as an example); generally, their sizes should be roughly equivalent. The distance between the magnet and the micro-magnetic cone is required to magnetize the micro-magnetic cone with an external field strength of 0.5T-0.7T (taking a microsphere diameter of 50 μm as an example) to form a micro-magnetic field. The width of the capture channel is related to the diameter of the magnetic microsphere, with a width within the range (3d, 4d] being optimal (d represents the diameter of the magnetic microsphere), ensuring that other uncaptured magnetic microspheres can pass through smoothly.

[0063] Example 3

[0064] This embodiment provides a detection system based on the biomolecular multiplex detection chip based on the coupling effect of microfluidic field and micromagnetic field described in Embodiment 1, such as... Figure 4 As shown, it includes a first reservoir that is connected to each of the first liquid inlets 102 and is used to store different types of functionalized magnetic microsphere solutions.

[0065] It is connected to the second liquid inlet 102 and is used as a second reservoir for the sample solution to be tested and the test solution;

[0066] The inlet is connected to the first outlet 20 and is used to store waste liquid in the first waste liquid pool 29;

[0067] A first negative pressure pump 30 is connected to the outlet of the first waste liquid tank 29;

[0068] The inlet is connected to the second outlet 13 and is used to store waste liquid in the second waste liquid pool 27;

[0069] A second negative pressure pump 28 connected to the outlet of the second waste liquid tank 27; and

[0070] A light source used to irradiate magnetic microspheres so that the presence of fluorescent magnetic microspheres can be determined by fluorescence signal detection.

[0071] Specifically, the first liquid reservoir includes a liquid reservoir 24 for storing a first type of functionalized magnetic microsphere solution, a liquid reservoir 25 for storing a second type of functionalized magnetic microsphere solution, and a liquid reservoir 26 for storing a third type of functionalized magnetic microsphere solution; wherein, the liquid reservoir 24 is connected to the liquid inlet 17, the liquid reservoir 25 is connected to the liquid inlet 18, and the liquid reservoir 26 is connected to the liquid inlet 19.

[0072] The second reservoir includes a reservoir 21 for storing a mixed solution of biomolecules, a reservoir 22 for storing fluorescently labeled detection reagents, and a reservoir 23 for storing eluent; wherein, the reservoir 21 is connected to the inlet 14, the reservoir 22 is connected to the inlet 15, and the reservoir 23 is connected to the inlet 16.

[0073] Furthermore, to achieve sample injection control of the functionalized magnetic microsphere solution, elastic valves 9, 10, 11, and 12 are provided on the lower substrate 4 of the chip. Among them, elastic valve 9 is located below the liquid inlet 19, elastic valve 10 is located below the liquid inlet 18, and elastic valve 11 is located below the liquid inlet 17, so as to control the opening and closing of the corresponding ball-catching channels.

[0074] The detection method of the detection system in this embodiment includes the following steps:

[0075] Step 1: Adjust the distance between magnet 2 and micro magnetic cones 6, 7 and 8 to a predetermined distance; so as to generate a primary magnetic field of 5500 Gauss to 7000 Gauss to magnetize the micro magnetic cones and form a micro magnetic field.

[0076] Step 2: Add the first type of functionalized magnetic microsphere liquid to the storage tank 24, open the elastic valve 11 and elastic valve 12, and then turn on the first negative pressure pump 30 so that the magnetic microspheres 31 entering the first capture chamber 53 are captured by the micro magnetic field formed by the micro magnetic cone 6, and then close the elastic valve 11.

[0077] Step 3: Add the second type of functionalized magnetic microsphere liquid to the storage tank 25, open the elastic valve 10 so that the magnetic microspheres 32 entering the second capture chamber 53 are captured by the micro magnetic field formed by the micro magnetic cone 7, and then close the elastic valve 10.

[0078] Step 4: Add the third functionalized magnetic microsphere liquid to the storage tank 26, open the elastic valve 9 so that the magnetic microspheres 33 entering the third capture chamber 53 are captured by the micro magnetic field formed by the micro magnetic cone 8, and then close the elastic valves 9 and 12 and the first negative pressure pump 30.

[0079] Of the steps above, steps 2, 3, and 4 can be performed simultaneously or separately.

[0080] Step 5: Add the biomolecule mixture to the storage tank 21, turn on the second negative pressure pump 28, and wait for a period of time until the biomolecule mixture and the magnetic microspheres are in full contact.

[0081] Step 6: Add the fluorescently labeled detection reagent to the storage tank 22 and wait for a period of time.

[0082] Step 7: Turn off the second negative pressure pump 28 and irradiate with laser.

[0083] Step 8: Observe whether magnetic microspheres 31, 32, and 33 produce fluorescence signals to obtain the detection results of whether the corresponding target analyte exists.

[0084] Step 9: Remove magnet 2 to release magnetic microspheres 31, 32, and 33; add eluent to the storage tank 23, open the elastic valve 12, and turn on the second negative pressure pump 28 to remove the magnetic microspheres from the chip. After cleaning, close the elastic valve 12 and the second negative pressure pump 28; the cleaned magnetic microspheres can be reused.

[0085] Step 10: Turn on the second negative pressure pump 28 to clean the residual mixed solution in the chip. After cleaning, turn off the second negative pressure pump 28.

[0086] In other embodiments, more ball-catching channels, capture cavities, and micro magnetic cones can be similarly arranged on a single chip substrate. Each capture cavity captures different types of magnetic microspheres. According to the above method, it is possible to simultaneously detect more types of biological samples on a single chip substrate.

[0087] The above description is merely a specific embodiment of this utility model, but the protection scope of this utility model is not limited thereto. Any variations or substitutions that can be easily conceived by those skilled in the art within the technical scope disclosed in this utility model should be included within the protection scope of this utility model. Therefore, the protection scope of this utility model should be determined by the protection scope of the claims.

Claims

1. A bio-molecular multiplex detection chip based on the coupling of micro-fluid field and micro-magnetic field, characterized in that, Including chip substrate; The top of the chip substrate is provided with a first liquid inlet, a second liquid inlet, a first liquid outlet, and a second liquid outlet; The chip substrate has a ball-catching channel, a detection channel, and a capture cavity inside; The first liquid inlet is configured as a plurality of the same number as the capture chamber, for correspondingly introducing different types of functionalized magnetic microsphere solutions; the second liquid inlet is used to introduce the sample solution to be tested and the detection solution. The first inlet of each capture chamber is connected to a first liquid inlet through the ball-catching channel, and the first outlet is connected to the first liquid outlet through the ball-catching channel. Each capture chamber is connected sequentially through the detection channel, wherein the second inlet of the first capture chamber is connected to the second liquid inlet through the detection channel, and the second outlet of the last capture chamber is connected to the second liquid outlet. The width of the ball-catching channel and the capturing cavity is greater than the diameter of the magnetic microspheres, and the width of the detection channel is less than the diameter of the magnetic microspheres; A micro magnetic cone is embedded in the chip substrate below the capture cavity to capture magnetic microspheres within the capture cavity.

2. The biomolecular multiplex detection chip based on the coupling effect of microfluidic field and micromagnetic field according to claim 1, characterized in that, The micro magnetic cone is a magnetic nanoparticle aggregated into a cone shape.

3. A biomolecular multiplex detection chip based on the coupling effect of microfluidic field and micromagnetic field according to any one of claims 1-2, characterized in that, The chip substrate includes an upper substrate and a lower substrate bonded together. The ball-catching channel, the detection channel, and the capture chamber are formed on the upper substrate, and the bottom wall is open. The micro magnetic cone is disposed on the lower substrate.

4. A biological molecule multiplex detection assembly based on the coupling of microfluidic field and micro-magnetic field, characterized in that, include: The biomolecular multiplex detection chip based on the coupling effect of microfluidic field and micromagnetic field as described in any one of claims 1-3; as well as A magnet is positioned below the biomolecular multiplex detection chip based on the coupling effect of microfluidic field and micromagnetic field, and the distance between the magnet and the micromagnetic cone can be adjusted.

5. The bio-molecular multiplex detection assembly based on micro-fluidic field and micro-magnetic field coupling effect according to claim 4, characterized in that, The magnet is a permanent magnet or an electromagnet.

6. The biomolecular multiplex detection component based on the coupling effect of microfluidic field and micromagnetic field according to claim 4 or 5, characterized in that, The width of the ball-catching channel ranges from (3d, 4d), where d represents the diameter of the magnetic microspheres.

7. A detection system based on the biomolecular multiplex detection component based on the coupling effect of microfluidic field and micromagnetic field as described in any one of claims 4-6, characterized in that, include: A first liquid reservoir is connected to each of the first liquid inlets and is used to store different types of functionalized magnetic microsphere solutions. A second reservoir connected to the second inlet and used for the sample solution to be tested and the detection solution; The inlet is connected to the first outlet and is used to store waste liquid in a first waste liquid tank. A first negative pressure pump connected to the outlet of the first waste liquid tank; The inlet is connected to the second outlet and is used to store waste liquid in a second waste liquid tank. A second negative pressure pump connected to the outlet of the second waste liquid tank; and A light source used to irradiate magnetic microspheres so that the presence of fluorescent magnetic microspheres can be determined by fluorescence signal detection.

8. The detection system according to claim 7, characterized in that: The bottom of the chip substrate is provided with elastic valves for opening and closing the first liquid inlet and the first liquid outlet, corresponding to the first liquid inlet and the first liquid outlet.

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