A micro-channel and its preparation method and application

Abstract

The present invention relates to a micro-channel and its preparation method and application. The micro-channel comprises: a linear hollow tube, with a first liquid inlet and a first liquid outlet respectively provided at both ends; the inner diameter of the linear hollow tube is 1-2 mm; the material of the linear hollow tube is silica gel; a first liquid inlet pipe, which is arranged on the tube wall of the linear hollow tube near one end of the reaction washing chamber and is connected to the reaction washing chamber; the material of the first liquid inlet pipe is silica gel; and, a first liquid outlet pipe, which is arranged on the tube wall of the linear hollow tube near the other end of the reaction washing chamber and is connected to the reaction washing chamber; the material of the first liquid outlet pipe is silica gel. How to provide a micro-channel, which realizes high-throughput reaction and separation through an integrated design of reaction, separation and washing, and also has a good washing effect; and the reaction liquid added to the reaction chamber can participate in the reaction, thereby improving the product yield and saving costs.

Description

Technical Field

[0001] The present invention belongs to the technical field of micro-pipes, and in particular relates to a micro-pipe and a preparation method and application thereof. Background Art

[0002] At present, the main structure of the existing microchannel for magnetic separation is a hollow tube, one end of which is provided with three inlets, which are used to slowly introduce reaction liquid (the reaction liquid includes the substance to be extracted and other impurities), magnetic microspheres (including the captured substance of the substance to be extracted) and water, and the other end is provided with an outlet; the inner diameter of the hollow tube is only a few hundred microns, the reaction liquid and water are in a layered state in the microchannel, and the magnetic microspheres are dispersed in the reaction liquid and water; a circular reaction chamber with a diameter of at least 1 mm is provided in the middle of the hollow tube for reaction and mixing of the reaction liquid, water and magnetic microspheres; during use, the permanent magnet is firstly controlled to translate manually or automatically to control the magnetic microspheres to move to the circular reaction chamber, and then the permanent magnet is manually or automatically controlled to rotate to rotate the magnetic microspheres in the circular reaction chamber, and under the rotating stirring of the magnetic microspheres, the substance to be extracted in the reaction liquid is combined with the magnetic microspheres, thereby realizing the separation of the substance to be extracted from other impurities; the microchannel ..., and the magnetic microspheres are dispersed in the Although the design of the pipeline can fully mix the magnetic microspheres with the reaction liquid and improve the separation efficiency, due to the design of the circular reaction chamber of the microchannel, it is difficult for the washing liquid to be flushed into the recessed area of ​​the circular reaction chamber, resulting in poor washing effect and low washing efficiency, and it is impossible to achieve online cleaning of the reaction product; and if the reaction area of ​​the microchannel is designed as a linear inner cavity with a tube diameter equal to that of other areas, although this design can be beneficial to washing, due to the limited space in the microchannel and the absence of a reaction chamber with a variable diameter, when the magnetic microspheres rotate in the microchannel, only the reaction liquid in the magnetic microsphere area can fully contact and react with the magnetic microspheres, while the reaction liquid outside the magnetic microsphere area is difficult to contact the magnetic microspheres, resulting in part of the reaction liquid being unable to contact and react with the captured substance of the substance to be extracted on the magnetic microspheres, thereby resulting in the inability to separate the part of the product, that is, the product cannot be fully collected, the product yield is low, and cost waste is caused. Summary of the invention

[0003] The main purpose of the present invention is to provide a microchannel and a preparation method and application thereof. The technical problem to be solved is how to provide a microchannel that realizes high-throughput reaction and separation through an integrated design of reaction, separation and washing, while also having a good washing effect; and the reaction liquid added to the reaction chamber can all participate in the reaction, thereby improving the product yield and saving costs.

[0004] The purpose of the present invention and the technical problem to be solved are achieved by adopting the following technical solutions. A micro-pipeline proposed in the present invention comprises:

[0005] A straight hollow tube, with a first liquid inlet and a first liquid outlet respectively disposed at both ends; the inner diameter of the straight hollow tube is 1 to 2 mm; the material of the straight hollow tube is silica gel; the middle section of the straight hollow tube is a reaction washing chamber;

[0006] a first liquid inlet pipe, which is arranged on the wall of the linear hollow tube near one end of the reaction and washing chamber and is connected to the reaction and washing chamber; the first liquid inlet pipe is used to inject reaction liquid or washing liquid into the reaction and washing chamber; the material of the first liquid inlet pipe is silica gel; and,

[0007] The first liquid outlet pipe is arranged on the wall of the linear hollow tube close to the other end of the reaction washing chamber and is connected with the reaction washing chamber; the first liquid outlet pipe is used to discharge reaction waste liquid; the material of the first liquid outlet pipe is silica gel.

[0008] Preferably, in the aforementioned micro-channel, a hydrophobic film is provided on the inner wall of the micro-channel, and the thickness of the hydrophobic film is 1 to 2 μm.

[0009] Preferably, in the aforementioned micro-channel, the first liquid inlet pipe and the first liquid outlet pipe are respectively arranged on two opposite sides of the tube wall of the linear hollow tube.

[0010] Preferably, in the aforementioned micro-channel, the first liquid inlet, the first liquid outlet, the first liquid inlet and the first liquid outlet are arranged in sequence along the axial direction of the linear hollow tube.

[0011] Preferably, in the aforementioned micro-channel, the linear hollow tube and the first liquid inlet tube are an integral structure; the linear hollow tube and the first liquid outlet tube are an integral structure.

[0012] Preferably, in the aforementioned microchannel, the silicone is polydimethylsiloxane.

[0013] The purpose of the present invention and the technical problem solved by the present invention are also achieved by adopting the following technical solutions. According to a method for preparing a micro-channel proposed by the present invention, the method comprises the following steps:

[0014] 1) Inject liquid silicone into the mold and solidify it;

[0015] 2) placing the solidified silica gel and the mold together in an organic solution to dissolve the mold to obtain microchannels; the solidified silica gel cannot be dissolved by the organic solution.

[0016] Preferably, in the aforementioned method for preparing microchannels, the organic solution is acetone; the material of the mold is acrylonitrile-butadiene-styrene; and the silicone is polydimethylsiloxane.

[0017] Preferably, in the aforementioned method for preparing the microchannel, a pulsed laser deposition method or a chemical vapor deposition method is used to plate a hydrophobic film on the inner wall of the microchannel; the hydrophobic film is made of polyparaxylene and has a thickness of 1 to 2 μm.

[0018] The purpose of the present invention and the technical problems solved therein are also achieved by adopting the following technical solutions: According to a magnetic separation device proposed by the present invention, the magnetic separation device comprises the above-mentioned micro-pipeline.

[0019] By means of the above technical solution, the micro-channel and its preparation method and application proposed by the present invention have at least the following advantages:

[0020] The inner diameter of the linear hollow tube in the microchannel described in the present invention is 1 to 2 mm, which is larger than the inner diameter of the conventional microchannel, so that it can achieve a large flux of injection of reaction liquid, water, magnetic beads and washing liquid, which can reach 1.6 mL / min. The inner diameter of the conventional microchannel is only a few hundred microns, and its liquid flux is only tens of microliters per minute. The design of the inner diameter size of the pipeline can improve the utilization efficiency of the equipment.

[0021] The micro-channel of the present invention is an important component of the magnetic separation device. The main structure of the micro-channel is a straight hollow tube with an inner diameter of 1 to 2 mm, with a first liquid inlet and a first liquid outlet at both ends, and the first liquid inlet pipe and the first liquid outlet are connected on the tube wall. The first liquid inlet pipe is used to inject reaction liquid or washing liquid, and the first liquid outlet pipe is used to discharge waste liquid. The area between the first liquid inlet pipe and the first liquid outlet pipe is a reaction washing chamber. This structural setting can make the internal space of the micro-channel tube larger (the inner diameter of the tube is 1 to 2 mm), and the reaction liquid, water, magnetic beads and washing liquid can be injected in large flux; on the other hand, the middle section of the micro-channel tube is set as a reaction washing chamber, and during the reaction and washing period, the reaction liquid, water, magnetic beads and washing liquid can be injected into the reaction washing chamber. , the magnetic microspheres can be made to stay in the reaction washing chamber by magnetic control, and can be translated in the reaction washing chamber under magnetic control; at the same time, the second liquid inlet pipe is directly connected to the reaction washing chamber, so that the reaction liquid or washing liquid can be directly introduced into the reaction washing chamber, while there is almost no reaction liquid in the non-reaction washing chamber part of the microchannel tube; when reacting, the reaction liquid is introduced into the reaction washing chamber, and the reaction liquid can fully contact and react with the captured substance of the substance to be extracted on the magnetic microspheres; after the reaction is completed, the washing liquid is introduced into it to clean the magnetic microspheres after capturing the reactants, thereby achieving high-throughput reaction and separation while having a good washing effect. Moreover, the reaction liquid is directly introduced into the reaction washing chamber, so that the reaction liquid can fully contact and react with the captured substance of the substance to be extracted on the magnetic microspheres, thereby increasing the proportion of reactant collection, high product yield, and cost saving.

[0022] The above description is only an overview of the technical solution of the present invention. In order to more clearly understand the technical means of the present invention and implement it according to the contents of the specification, the following is a detailed description of the preferred embodiments of the present invention in conjunction with the accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 It is a schematic diagram of the structure of the micro-pipe of the present invention;

[0024] Figure 2 This is a schematic diagram of the overall structure of the magnetic separation device according to an embodiment of the present invention;

[0025] Figure 3 A schematic diagram of the structure and reaction process of the micro-channel according to an embodiment of the present invention;

[0026] Figure 4 A schematic diagram of a driving circuit in a magnetic separation device according to an embodiment of the present invention;

[0027] Figure 5 This is a schematic diagram of the dimensions of the micro-pipeline of the present invention;

[0028] Figure 6 A pulse timing diagram of the annular magnetic coil according to an embodiment of the present invention;

[0029] Figure 7 Schematic diagram of the preparation steps of the micro-channel according to an embodiment of the present invention;

[0030] Figure 8 Schematic diagram of the structure of the micro-pipeline of the present invention. DETAILED DESCRIPTION

[0031] In order to further explain the technical means and effects adopted by the present invention to achieve the predetermined invention purpose, the following is a detailed description of a micro-channel and its preparation method and application proposed by the present invention, its specific implementation method, structure, characteristics and effects in combination with the accompanying drawings and preferred embodiments. In the following description, different "one embodiment" or "embodiment" does not necessarily refer to the same embodiment. In addition, specific features, structures or characteristics in one or more embodiments may be combined in any suitable form.

[0032] The present invention proposes a micro-channel, as shown in the attached Figure 1 Said, it includes:

[0033] A straight hollow tube, with a first liquid inlet and a first liquid outlet respectively disposed at both ends; the inner diameter of the straight hollow tube is 1 to 2 mm; the material of the straight hollow tube is silica gel; the middle section of the straight hollow tube is a reaction washing chamber;

[0034] a first liquid inlet pipe, which is arranged on the wall of the linear hollow tube near one end of the reaction and washing chamber and is connected to the reaction and washing chamber; the first liquid inlet pipe is used to inject reaction liquid into the linear hollow tube; the material of the first liquid inlet pipe is silica gel; and,

[0035] The first liquid outlet pipe is arranged on the wall of the linear hollow tube close to the other end of the reaction washing chamber and is connected to the inner cavity of the linear hollow tube connected to the reaction washing chamber; the first liquid outlet pipe is used to discharge reaction waste liquid; the material of the first liquid outlet pipe is silica gel.

[0036] The micro-channel of the present invention is an important component of the magnetic separation device. The main structure of the micro-channel is a straight hollow tube with an inner diameter of 1 to 2 mm and an outer diameter of 2.4 to 4.5 mm. A first liquid inlet and a first liquid outlet are provided at both ends. The first liquid inlet pipe and the first liquid outlet pipe are connected on the tube wall of the straight hollow tube. The first liquid inlet pipe is used to inject the reaction liquid into the straight hollow tube, and the first liquid outlet pipe is used to discharge the liquid in the straight hollow tube. Figure 1 As shown, although the microchannel designed in the present invention is a straight hollow tube, a reaction washing chamber 105 is set between the first liquid inlet pipe 103 and the first liquid outlet pipe 104. During use, a plug or other tool is used to block the first liquid inlet and the first liquid outlet, and then the reaction liquid is injected. After the reaction liquid is injected, most of the reaction liquid exists in the reaction washing chamber 105, and the liquid inlet chamber 106 and the liquid outlet chamber 107 contain only a very small amount of reaction liquid (negligible). Therefore, during the separation process, the magnetic beads can fully react with the reaction liquid, thereby fixing the target in the reaction liquid on the magnetic beads, thereby improving the product yield and saving costs.

[0037] The present invention connects the first liquid inlet pipe and the first liquid outlet pipe with the linear hollow pipe. The connection means that the liquid can flow from the first liquid inlet pipe or the first liquid outlet pipe into the linear hollow pipe, or flow out from the linear hollow pipe into the first liquid inlet pipe or the first liquid outlet pipe. The connection between the first liquid inlet pipe and the first liquid outlet pipe and the linear hollow pipe is smooth without any obstruction, so that the reaction liquid and / or magnetic beads can flow in or out smoothly, avoiding accumulation at the connection of the magnetic beads, resulting in pipe blockage, thereby ensuring the normal use of the micro-pipe.

[0038] The microchannel prepared by the present invention needs to be placed in a magnetic field to be used normally. The first liquid inlet and the first liquid outlet can be used as the inlet or outlet of the sample solution. The microchannel of the present invention can also be used in the following method, such as Figure 8 As shown, the sample enters from the first liquid outlet 102, the product is discharged from the first liquid inlet 101, the reaction liquid is injected from the first liquid inlet pipe 103, and the waste liquid is discharged from the first liquid outlet pipe 104. Before operation, it is necessary to ensure that the microchannel is filled with solvent (the solvent is usually water).

[0039] The difference between the inner diameter of the first liquid inlet tube and the first liquid outlet tube of the present invention and the inner diameter of the linear hollow tube is -0.2 to 0.2 mm. For example, when the inner diameter of the linear hollow tube is 1 mm, the inner diameter of the first liquid inlet tube is between 0.8 and 1.2 mm, and the inner diameter of the first liquid outlet tube is also between 0.8 and 1.2 mm. Preferably, the inner diameters of the first liquid inlet tube, the first liquid outlet tube and the linear hollow tube are the same. By controlling the diameters of the linear hollow tube, the first liquid inlet tube and the first liquid outlet tube, a large-throughput injection of reaction liquid, water, magnetic beads and washing liquid can be achieved, which can reach 1.6 mL / min, while conventional equipment only has tens of microliters per minute, thereby improving the use efficiency of the equipment.

[0040] Preferably, in the aforementioned micro-channel, a hydrophobic film is provided on the inner wall of the micro-channel, and the thickness of the hydrophobic film is 1 to 2 μm.

[0041] Preferably, in the aforementioned micro-channel, the first liquid inlet pipe and the first liquid outlet pipe are respectively arranged on two opposite sides of the tube wall of the linear hollow tube.

[0042] Preferably, in the aforementioned micro-channel, the first liquid inlet, the first liquid outlet, the first liquid inlet and the first liquid outlet are arranged in sequence along the axial direction of the linear hollow tube.

[0043] The first liquid inlet pipe and the first liquid outlet pipe of the present invention can be arranged on the same side or on both sides of the linear hollow tube 1, wherein Figure 1 The setting is the best choice, that is, the first liquid inlet pipe 103 and the first liquid outlet pipe 104 are distributed on both sides of the straight hollow tube 1, and along the axial direction of the straight hollow tube, the first liquid inlet, the first liquid outlet, the first liquid inlet and the first liquid outlet are arranged in sequence; when in use, Figure 1 As shown, a pipette is used to inject a sample (the sample includes a solvent and magnetic beads) from the first liquid inlet 101 into the linear hollow tube 1, and then under the action of an external magnetic field, the magnetic beads move to the reaction washing chamber 105 and mix with the reaction solution injected by the first liquid inlet pipe 103. By controlling the magnetic field, the magnetic beads are made to reciprocate in the reaction washing chamber, so that more target objects in the reaction solution are fixed on the magnetic beads; after the target objects in the reaction solution are fixed on the magnetic beads, the first liquid inlet pipe 103 stops introducing the reaction solution and introduces washing water instead to start washing. The washing process is the same as the reaction separation process. By controlling the magnetic field, the magnetic beads fixed with the target objects are made to reciprocate in the reaction washing chamber, so as to achieve the washing effect; after the washing is completed, the product containing the magnetic beads and the target object is discharged from the first liquid outlet 102, and the waste liquid such as the reaction solution and the washing water is discharged from the first liquid outlet pipe, so that the reaction, separation and washing are integrated in the same container, which can avoid product loss and improve process efficiency.

[0044] also, Figure 8The method of using can also be used as a means of using microchannels, as long as the flow direction of the reaction solution in the microchannel is opposite to the flow direction of the magnetic beads, such as Figure 3 As shown, the flow direction of the reaction liquid in the microchannel refers to the flow direction of the reaction liquid in the reaction washing chamber. At the beginning of the reaction, this direction is opposite to the flow direction of the magnetic beads; when the reaction starts, the magnetic beads reciprocate in the reaction washing chamber.

[0045] like Figure 3 As shown, the straight hollow tube between the first liquid inlet tube 103 and the first liquid outlet tube 104 of the present invention is a reaction washing chamber. This arrangement can prevent the reaction liquid from flowing into the liquid inlet chamber and the liquid outlet chamber, thereby preventing the magnetic beads from being unable to contact all of the reaction liquid, thereby causing part of the target in the reaction liquid to not be extracted, resulting in waste of the reaction liquid; on the other hand, the reaction liquid in the reaction washing chamber flows from right to left, while the magnetic beads flow from left to right. Through the counter-flow of the reaction liquid and the magnetic beads, the magnetic beads can contact more reaction liquid, thereby improving the extraction efficiency of the target.

[0046] Preferably, in the aforementioned micro-channel, the linear hollow tube and the first liquid inlet tube are an integral structure; the linear hollow tube and the first liquid outlet tube are an integral structure. The integral structure means that the linear hollow tube is integrally formed during the preparation process, and there is no connector at the connection between the linear hollow tube and the first liquid inlet tube and the first liquid outlet tube, and it is not removable. The integrally formed structural design, combined with the silicone material, makes the micro-channel more convenient to disassemble.

[0047] Preferably, in the aforementioned microchannel, the silicone is polydimethylsiloxane.

[0048] Since the microchannel needs to be used in a magnetic field environment, the material of the microchannel needs to be a material that cannot be magnetized. The present invention chooses to use polydimethylsiloxane to prepare the microchannel, because polydimethylsiloxane is softer than materials such as ceramics and glass, can be bent, and is easy to disassemble and clean; but due to the characteristics of polydimethylsiloxane itself, it will adsorb magnetic beads, targets and other substances. Therefore, the present invention coats a hydrophobic film on the inner wall of the microchannel. The thickness of the hydrophobic film can ensure that the microchannel does not adsorb the sample in the microchannel. If it is too thick, it will cause a waste of space inside the microchannel. The thickness of the hydrophobic film is preferably 1-2μm, and the material of the hydrophobic film is polyparaxylene film.

[0049] The present invention also provides a method for preparing a micro-channel, which comprises the following steps:

[0050] 1) Inject liquid silicone into the mold and solidify it;

[0051] 2) placing the solidified silica gel and the mold together in an organic solution to dissolve the mold to obtain microchannels; the solidified silica gel cannot be dissolved by the organic solution.

[0052] Preferably, in the aforementioned method for preparing microchannels, the organic solution is acetone; the material of the mold is acrylonitrile-butadiene-styrene; and the silicone is polydimethylsiloxane.

[0053] The mold can be purchased or prepared, as long as the structure of the prepared micro-pipe meets the requirements. The preparation method can adopt 3D printing and other methods. When preparing the micro-pipe, the present invention adopts the method of dissolving the mold with an organic solution to prepare the micro-pipe. This preparation method can prevent the micro-pipe from being damaged during demolding, resulting in the micro-pipe being unusable. In the preparation process, the solidified silica gel is placed in an organic solution together with the mold, so the selection of the material of the mold, the material of the micro-pipe and the organic solvent is particularly important. The organic solution of the present invention is acetone, the material of the mold is acrylonitrile-butadiene-styrene, and the silica gel is polydimethylsiloxane. The selection of the organic solvent and the mold can also be selected from other substances, as long as the organic solution can dissolve the mold and the micro-pipe is not dissolved by the organic solvent.

[0054] Preferably, in the aforementioned method for preparing the microchannel, a pulsed laser deposition method or a chemical vapor deposition method is used to plate a hydrophobic film on the inner wall of the microchannel; the hydrophobic film is made of polyparaxylene and has a thickness of 1 to 2 μm.

[0055] The present invention also provides a magnetic separation device, which includes the above-mentioned micro-channel.

[0056] like Figures 2-3 As shown, the magnetic separation device comprises:

[0057] Micro-channel 11, which is the above-mentioned micro-channel;

[0058] The coil array 2 comprises a plurality of annular magnetic coils 21 arranged in parallel; the micro-pipeline 11 is arranged to penetrate the inner cavity of the annular magnetic coil; the annular magnetic coils are arranged at intervals on the outside of the micro-pipeline; the outer diameter of the annular magnetic coil is 8 to 15 mm, and the thickness is 2 to 5 mm; the interval between adjacent annular magnetic coils is 2 to 4 mm;

[0059] When the micro-channel of the present invention is used in a magnetic separation device, its size needs to be adjusted. In the magnetic separation device, the coil array is a device that provides a magnetic field environment. In order to achieve the purpose of controlling the magnetic beads in the micro-channel by adjusting the magnetic field, the present invention sets the spacing between adjacent annular magnetic coils in the coil array to 2 to 4 mm. Therefore, when selecting the length of the micro-channel, it is necessary to make the liquid inlet chamber, the reaction washing chamber, and the liquid outlet chamber able to accommodate a corresponding number of electromagnetic coils. Preferably, the size of the micro-channel is as follows: Figure 5 As shown, Figure 5 The widths of the first liquid outlet 102 and the first liquid outlet pipe 104 are both outer diameters.

[0060] The magnetic beads 200 include magnetic microspheres 201 and magnetic particles 202; the magnetic beads 200 can move in the microchannel 11 under the action of the plurality of annular magnetic coils 21; the magnetic microspheres 201 are magnetic metals with an organic layer coated on the surface; the magnetic particles 202 are magnetic metals without a coating layer, the particle size of the magnetic microspheres 201 is 10 to 50 μm, and the particle size of the magnetic particles 202 is less than 5 μm; the mass ratio of the magnetic microspheres 201 to the magnetic particles 202 is 1:0.2 to 1.2;

[0061] The magnetic beads are composed of magnetic microspheres and magnetic particles. The magnetic microspheres are loaded with a first substance, which can react with a second substance (target substance) in a reaction solution, so that the target substance is fixed on the magnetic microspheres, thereby realizing separation of the target substance from other substances in the reaction solution. The present invention controls the particle size of the magnetic microspheres and the magnetic particles, and the mass ratio of the magnetic microspheres and the magnetic particles, so that the magnetic beads can be driven by a weak magnetic field, and then smaller experimental equipment can be used, thereby realizing miniaturization of the device.

[0062] A fixed circuit board 4 is provided with a pin row at one end of its surface, and the pin row is inserted into the socket to electrically connect the fixed circuit board with the driving circuit board 3; two rows of conductive holes 41 are provided on the surface of the fixed circuit board, with 14 conductive holes in each row, for a total of 28 conductive holes; through holes are provided at the four corners of the fixed circuit board 4 and the driving circuit board 3, and M2 screws are used to fix the fixed circuit board 4 and the driving circuit board 3;

[0063] The main function of the fixing circuit is to fix the coil array and electrically connect the coil array to the driving circuit, so as to control the coil array through the driving circuit and thus achieve the purpose of controlling the magnetic field.

[0064] Drive circuit board 3, such as Figure 4As shown, it includes a boost converter 36 electrically connected to 7 magnetic field generator drivers 34; the magnetic field generator driver shown is a ring-shaped magnetic coil driver; the ring-shaped magnetic coil drivers are connected in parallel with each other; the voltage is adjusted from 5V to 12V; the processor 32 is a micro control unit MICRO CONTROL UNIT processor, which is electrically connected to the 7 ring-shaped magnetic coil drivers through 7 NMOS field effect transistors; the NMOS field effect transistors correspond to the ring-shaped magnetic coil drivers one by one and are connected in series; the NMOS is an N-Metal-Oxide-Semiconductor; each of the ring-shaped magnetic coil drivers is connected in parallel with a Schottky diode 35; the data line interface 31 is a universal serial bus UNIVERSAL SERIAL BUS interface, which is connected to the user end 10 through a data line 111; and,

[0065] A socket, which is arranged at one end of the surface of the driving circuit board 3;

[0066] The lower positioner 5 includes a plurality of first grooves 51 arranged in parallel; the annular magnetic coil 21 is inserted into the first grooves 51 to fix the position of the annular magnetic coil 21; the two ends of the lower positioner 5 also include second grooves 52; the two ends of the micro-pipe 11 are fixedly connected to the second grooves 52;

[0067] The upper positioner 6 includes a plurality of third grooves 61 arranged in parallel; the third grooves 61 correspond to the first grooves 51 one by one and are used to fix the position of the annular magnetic coil 21; the two ends of the upper positioner 6 also include fourth grooves 62; the fourth grooves 62 are clamped at the two ends of the micro-pipe 11 to fix the position of the micro-pipe 11;

[0068] A base 8, used to support the entire device;

[0069] The first groove 51 can fix the position of the annular magnetic coil 21, so that the spacing of the annular magnetic coil 21 is fixed, and prevent the spacing of the annular magnetic coil from changing due to human error or other situations, so that the control of the magnetic beads is affected. The second groove 52 is set at both ends of the lower positioner, which can support the micro-pipeline, so that the micro-pipeline can be placed horizontally and pass through the annular magnetic coil;

[0070] The third groove 61 cooperates with the first groove 51 to fix the position of the annular magnetic coil so that the spacing distance of the annular magnetic coil does not change. The fourth groove 62 fixes both sides of the micro-pipe 11 from both sides to prevent the micro-pipe 11 from lateral displacement.

[0071] The method of using the magnetic separation equipment is as follows: Figure 3For example: the magnetic beads 200 loaded with the first substance 2011 enter the liquid inlet chamber 106 under the action of the annular magnetic coils 211 and 212; then enter the reaction washing chamber 105 under the action of the annular magnetic coils 213, 214 and 215, and the reaction liquid is introduced through the first liquid inlet pipe 103. Under the action of the annular magnetic coils 213, 214 and 215, the magnetic beads reciprocate in the reaction washing chamber 105, and are fully mixed and reacted with the second substance 2012 in the reaction liquid; after the reaction is completed, the reaction liquid is stopped and washing water is introduced instead, and the magnetic beads still reciprocate in the reaction washing chamber 105, thereby achieving full washing; after washing, under the action of the annular magnetic coils 216 and 217, the magnetic beads are discharged from the first liquid outlet 102 through the liquid outlet chamber 107 to obtain a product; the waste liquid is discharged from the first liquid outlet pipe 104. In addition, the device has other methods of use, for example: magnetic beads can also enter from the first liquid outlet 102 and be discharged from the first liquid inlet 101, reaction liquid or washing water is injected from the first liquid outlet pipe 104 and discharged from the first liquid inlet pipe 103.

[0072] The reaction described in the present invention refers to a reaction in which the first substance loaded on the magnetic beads can bind to the second substance (target substance) in the reaction solution. The binding method can also be gene complementation or other means, as long as the second substance can be fixed on the magnetic beads.

[0073] The present invention will be further described below in conjunction with specific embodiments, but this should not be construed as limiting the scope of protection of the present invention. Some non-essential improvements and adjustments made to the present invention by technicians in this field based on the above-mentioned contents of the present invention still fall within the scope of protection of the present invention.

[0074] Unless otherwise specified, the materials, reagents, etc. involved below are all commercially available products well known to those skilled in the art; unless otherwise specified, the methods described are all methods well known in the art. Unless otherwise defined, the technical terms or scientific terms used should have the common meanings understood by ordinary technicians in the field to which the present invention belongs.

[0075] Example 1

[0076] A micro-channel comprising:

[0077] A straight hollow tube, with a first liquid inlet and a first liquid outlet at both ends thereof; the outer diameter of the straight hollow tube is 2.4 mm, and the inner diameter is 1 mm; the material of the straight hollow tube is polydimethylsiloxane; the middle section of the straight hollow tube is a reaction washing chamber;

[0078] a first liquid inlet pipe, which is arranged on the wall of the linear hollow tube close to one end of the reaction and washing chamber and communicates with the inner cavity of the linear hollow tube; the first liquid inlet pipe is used to inject reaction liquid or washing liquid into the reaction and washing chamber; the material of the first liquid inlet pipe is polydimethylsiloxane; and,

[0079] A first liquid outlet pipe, which is arranged on the wall of the linear hollow tube close to the other end of the reaction and washing chamber and communicates with the reaction and washing chamber to form an inner cavity of the linear hollow tube; the first liquid outlet pipe is used to discharge reaction waste liquid; the material of the first liquid outlet pipe is polydimethylsiloxane;

[0080] A hydrophobic film is provided on the inner wall of the microchannel, and the thickness of the hydrophobic film is 1 μm;

[0081] The first liquid inlet pipe and the first liquid outlet pipe are respectively arranged on opposite sides of the tube wall of the linear hollow tube; along the axial direction of the linear hollow tube, the first liquid inlet, the first liquid outlet pipe, the first liquid inlet pipe and the first liquid outlet are arranged in sequence; the linear hollow tube and the first liquid inlet pipe are an integral structure; the linear hollow tube and the first liquid outlet pipe are an integral structure.

[0082] Example 2

[0083] A micro-channel comprising:

[0084] A straight hollow tube, with a first liquid inlet and a first liquid outlet at both ends thereof; the outer diameter of the straight hollow tube is 3 mm, and the inner diameter is 1.5 mm; the material of the straight hollow tube is polydimethylsiloxane; the middle section of the straight hollow tube is a reaction washing chamber;

[0085] a first liquid inlet pipe, which is arranged on the wall of the linear hollow tube close to one end of the reaction and washing chamber and communicates with the inner cavity of the linear hollow tube; the first liquid inlet pipe is used to inject reaction liquid or washing liquid into the reaction and washing chamber; the material of the first liquid inlet pipe is polydimethylsiloxane; and,

[0086] A first liquid outlet pipe, which is arranged on the wall of the linear hollow tube close to the other end of the reaction and washing chamber and communicates with the reaction and washing chamber to form an inner cavity of the linear hollow tube; the first liquid outlet pipe is used to discharge reaction waste liquid; the material of the first liquid outlet pipe is polydimethylsiloxane;

[0087] A hydrophobic film is provided on the inner wall of the microchannel, and the thickness of the hydrophobic film is 1.5 μm;

[0088] The first liquid inlet pipe and the first liquid outlet pipe are respectively arranged on opposite sides of the tube wall of the linear hollow tube; along the axial direction of the linear hollow tube, the first liquid inlet, the first liquid outlet pipe, the first liquid inlet pipe and the first liquid outlet are arranged in sequence; the linear hollow tube and the first liquid inlet pipe are an integral structure; the linear hollow tube and the first liquid outlet pipe are an integral structure.

[0089] Example 3

[0090] A micro-channel comprising:

[0091] A straight hollow tube, with a first liquid inlet and a first liquid outlet at both ends thereof; the outer diameter of the straight hollow tube is 4.5 mm, and the inner diameter is 2 mm; the material of the straight hollow tube is polydimethylsiloxane; the middle section of the straight hollow tube is a reaction washing chamber;

[0092] a first liquid inlet pipe, which is arranged on the wall of the linear hollow tube close to one end of the reaction and washing chamber and communicates with the inner cavity of the linear hollow tube; the first liquid inlet pipe is used to inject reaction liquid or washing liquid into the reaction and washing chamber; the material of the first liquid inlet pipe is polydimethylsiloxane; and,

[0093] A first liquid outlet pipe, which is arranged on the wall of the linear hollow tube close to the other end of the reaction and washing chamber and communicates with the reaction and washing chamber to form an inner cavity of the linear hollow tube; the first liquid outlet pipe is used to discharge reaction waste liquid; the material of the first liquid outlet pipe is polydimethylsiloxane;

[0094] A hydrophobic film is provided on the inner wall of the microchannel, and the thickness of the hydrophobic film is 2 μm;

[0095] The first liquid inlet pipe and the first liquid outlet pipe are respectively arranged on opposite sides of the tube wall of the linear hollow tube; along the axial direction of the linear hollow tube, the first liquid inlet, the first liquid outlet pipe, the first liquid inlet pipe and the first liquid outlet are arranged in sequence; the linear hollow tube and the first liquid inlet pipe are an integral structure; the linear hollow tube and the first liquid outlet pipe are an integral structure.

[0096] Example 4

[0097] A micro-channel comprising:

[0098] A straight hollow tube, with a first liquid inlet and a first liquid outlet at both ends thereof; the outer diameter of the straight hollow tube is 2.5 mm, and the inner diameter is 1 mm; the material of the straight hollow tube is polydimethylsiloxane; the middle section of the straight hollow tube is a reaction washing chamber;

[0099] a first liquid inlet pipe, which is arranged on the wall of the linear hollow tube close to one end of the reaction and washing chamber and communicates with the inner cavity of the linear hollow tube; the first liquid inlet pipe is used to inject reaction liquid or washing liquid into the reaction and washing chamber; the material of the first liquid inlet pipe is polydimethylsiloxane; and,

[0100] A first liquid outlet pipe, which is arranged on the wall of the linear hollow tube close to the other end of the reaction and washing chamber and communicates with the reaction and washing chamber to form an inner cavity of the linear hollow tube; the first liquid outlet pipe is used to discharge reaction waste liquid; the material of the first liquid outlet pipe is polydimethylsiloxane;

[0101] A hydrophobic film is provided on the inner wall of the microchannel, and the thickness of the hydrophobic film is 1 μm;

[0102] The first liquid inlet pipe and the first liquid outlet pipe are respectively arranged on opposite sides of the tube wall of the linear hollow tube; along the axial direction of the linear hollow tube, the first liquid inlet, the first liquid outlet pipe, the first liquid inlet pipe and the first liquid outlet are arranged in sequence; the linear hollow tube and the first liquid inlet pipe are an integral structure; the linear hollow tube and the first liquid outlet pipe are an integral structure.

[0103] Example 5

[0104] A method for preparing a microchannel, such as Figure 7 As shown, it includes the following steps:

[0105] 1) preparing a mold by 3D printing, injecting liquid polydimethylsiloxane into the mold, and solidifying it;

[0106] 2) placing the solidified polydimethylsiloxane and the mold in acetone to dissolve the mold to obtain microchannels; the material of the mold is acrylonitrile-butadiene-styrene;

[0107] A pulsed laser deposition method is used to plate a hydrophobic film on the inner wall of the micro-channel; the hydrophobic film is made of polyparaxylene and has a thickness of 1 μm.

[0108] Example 6

[0109] The present invention provides an integrated magnetic separation device, such as Figures 2 to 4 As shown, it includes:

[0110] The micro-channel has the same structure as that of Example 4 and a length of 49 mm;

[0111] The magnetic field module includes 7 annular magnetic coils arranged in parallel; the 7 annular magnetic coils are arranged with the central axis; the micro-pipe 11 passes through the annular magnetic coils along the central axis; the interval between adjacent annular magnetic coils is 3mm; the inner diameter of the annular magnetic coil 21 is 4mm, the outer diameter is 10mm, the thickness is 3mm, and the number of turns is 300.

[0112] Magnetic beads 200 include magnetic microspheres 201 and magnetic particles 202; the magnetic microspheres 201 are magnetic metals with an organic layer coated on the surface; the magnetic particles 202 are magnetic metals without a coating layer, the particle size of the magnetic microspheres 201 is 20 μm, and the particle size of the magnetic particles 202 is 1 μm; the mass ratio of the magnetic microspheres 201 to the magnetic particles 202 is 1:1.

[0113] A fixed circuit board 4 is provided with a pin row 42 at one end of its surface, and the pin row is inserted into the power strip to electrically connect the fixed circuit board with the driving circuit board 3; two rows of conductive holes 41 are provided on the surface of the fixed circuit board, with 14 conductive holes in each row, a total of 28 conductive holes, and the annular magnetic coil 21 is energized through the magnetic coil connector 7; through holes are provided at the four corners of the fixed circuit board 4 and the driving circuit board 3, and the fixed circuit board 4 and the driving circuit board 3 are fixedly connected using M2 screws;

[0114] A driving circuit board 3, which includes a boost converter 36 electrically connected to 7 magnetic field generator drivers; the magnetic field generator driver shown is a ring-shaped magnetic coil driver; the ring-shaped magnetic coil drivers are connected in parallel with each other; the voltage is adjusted from 5V to 12V; the processor 32 is a micro control unit MICRO CONTROL UNIT processor, which is electrically connected to the 7 ring-shaped magnetic coil drivers through 7 NMOS field effect transistors; the NMOS field effect transistors correspond to the ring-shaped magnetic coil drivers one by one and are connected in series; the NMOS is an N-Metal-Oxide-Semiconductor; each of the ring-shaped magnetic coil drivers is connected in parallel with a Schottky diode; the data line interface is a universal serial bus UNIVERSALSERIAL BUS interface, which is connected to the user end 10 through a data line 11; and,

[0115] The power strip is arranged at one end of the surface of the driving circuit board 3 .

[0116] The supporting module comprises: a lower positioner 5, which comprises 7 first grooves 51 arranged in parallel; the annular magnetic coil 21 is inserted into the first grooves 51 to fix the position of the annular magnetic coil 21; the two ends of the lower positioner 5 also comprise second grooves 52; the two ends of the micro-pipe 11 are fixedly connected to the second grooves 52;

[0117] The upper positioner 6 includes 7 third grooves 61 arranged in parallel; the third grooves 61 correspond to the first grooves 51 one by one and are used to fix the position of the annular magnetic coil 21; the two ends of the upper positioner 6 also include fourth grooves 62; the fourth grooves 62 are clamped at the two ends of the micro-pipe 11 to fix the position of the micro-pipe 11; and,

[0118] The usage is as follows: Figure 3 As shown:

[0119] S1, using a pipette to inject a sample from the first liquid inlet 101 of the microchannel 11; the sample includes: water, carboxylated magnetic microspheres loaded with DNA samples, and magnetic particles;

[0120] S2, start the magnetic coils 211 and 212 disposed outside the liquid inlet chamber 106 to introduce the sample into the liquid inlet chamber 106;

[0121] S3, starting the magnetic coils 213, 214 and 215 disposed outside the reaction and washing chamber 105 to introduce the sample into the reaction and washing chamber 105;

[0122] S4, using a syringe pump to inject the reaction liquid into the microchannel 11 through the first liquid inlet pipe 103, and then the magnetic coils 213, 214 and 215 work in a time sequence pulse to react; using a waste liquid collector to collect the waste liquid discharged from the first liquid outlet pipe 104;

[0123] S5, after the reaction is completed, stop introducing the reaction solution, use a syringe pump to inject washing solution into the microchannel 11 through the first liquid inlet pipe 103, and then use the magnetic coils 213, 214 and 215 to work in a time sequence pulse to wash; use a waste liquid collector to collect the waste liquid discharged from the first liquid outlet pipe 104;

[0124] S6. Use the first liquid outlet 102 of the finished product collector to collect the finished product 203.

[0125] The above steps can be performed in the PC client program in automatic mode or manual mode, as follows:

[0126] In the automatic mode, according to the above steps S1 to S6, the magnetic coil is controlled according to a predetermined time sequence, such as Figure 6 As shown, Figure 6 The horizontal axis is time t, and the vertical axis is the pulses of multiple magnetic coils. The steps are as follows:

[0127] In the sample injection time period T0, magnetic coil 1 (i.e. 211) and magnetic coil 2 (i.e. 212) work according to the time sequence pulse; in the reaction or washing time period T1, magnetic coil (i.e. 213), magnetic coil (i.e. 214), and magnetic coil 5 (i.e. 215) work according to the time sequence pulse;

[0128] During the product collection time period T2, the magnetic coil 6 (ie, 216) and the magnetic coil 7 (ie, 217) operate in a time sequence pulse manner.

[0129] In manual mode, the user can also independently control the on or off state of each magnetic coil to support manual operation to cooperate with the implementation of steps S1 to S6. After the reaction, the above step S5 can also be performed separately to wash the magnetic beads inside the microchannel.

[0130] In the above embodiments, the description of each embodiment has its own emphasis. For parts that are not described in detail in a certain embodiment, reference can be made to the relevant descriptions of other embodiments.

[0131] The above description is only a preferred embodiment of the present invention and does not constitute any form of limitation to the present invention. Although the present invention has been disclosed as a preferred embodiment as above, it is not intended to limit the present invention. Any technician familiar with the profession can make some changes or modifications to equivalent embodiments of equivalent changes using the technical contents disclosed above without departing from the scope of the technical solution of the present invention. However, any simple modification, equivalent change and modification made to the above embodiments based on the technical essence of the present invention without departing from the content of the technical solution of the present invention still falls within the scope of the technical solution of the present invention.

Claims

1. A micro-channel, characterized in that: It includes: A straight hollow tube, with a first liquid inlet and a first liquid outlet respectively disposed at both ends; the inner diameter of the straight hollow tube is 1 to 2 mm; The material of the straight hollow tube is silica gel; the middle section of the straight hollow tube is a reaction washing chamber; a first liquid inlet pipe, which is arranged on the wall of the linear hollow tube near one end of the reaction and washing chamber and is connected to the reaction and washing chamber; the first liquid inlet pipe is used to inject reaction liquid or washing liquid into the reaction and washing chamber; the material of the first liquid inlet pipe is silica gel; and, A first liquid outlet pipe, which is arranged on the wall of the linear hollow tube near the other end of the reaction and washing chamber and is connected to the reaction and washing chamber; the first liquid outlet pipe is used to discharge the reaction waste liquid; the material of the first liquid outlet pipe is silica gel; The first liquid inlet pipe and the first liquid outlet pipe are respectively arranged on two opposite sides of the tube wall of the linear hollow tube; The micro-channel is used by controlling the annular magnetic coil; the micro-channel runs through the inner cavity of the annular magnetic coil; the annular magnetic coil is arranged at intervals on the outer side of the micro-channel; the magnetic beads are controlled to reciprocate in the reaction washing chamber by the annular magnetic coil; The magnetic beads are composed of magnetic microspheres and magnetic particles.

2. The micro-duct according to claim 1, characterized in that: A hydrophobic film is arranged on the inner wall of the microchannel, and the thickness of the hydrophobic film is 1-2 μm.

3. The micro-duct according to claim 1, characterized in that: Along the axial direction of the linear hollow tube, the first liquid inlet, the first liquid outlet, the first liquid inlet and the first liquid outlet are arranged in sequence.

4. The micro-duct according to claim 1, characterized in that: The straight hollow tube and the first liquid inlet tube are an integral structure; the straight hollow tube and the first liquid outlet tube are an integral structure.

5. The micro-duct according to claim 1, characterized in that: The silica gel is polydimethylsiloxane.

6. A method for preparing a micro-channel according to claim 1, characterized in that: It includes the following steps: 1) Inject liquid silicone into the mold and solidify it; 2) placing the solidified silicone together with the mold in an organic solution to dissolve the mold and obtain microchannels; the solidified silicone cannot be dissolved by the organic solution.

7. The method for preparing a micro-channel according to claim 6, characterized in that: The organic solution is acetone; the material of the mold is acrylonitrile-butadiene-styrene; and the silicone is polydimethylsiloxane.

8. The method for preparing a micro-channel according to claim 6, characterized in that: A pulse laser deposition method or a chemical vapor deposition method is adopted to plate a hydrophobic film on the inner wall of the micro-channel; the hydrophobic film is made of polyparaxylene and has a thickness of 1 to 2 μm.

9. A magnetic separation device, characterized in that: The magnetic separation device comprises the micro-channel according to any one of claims 1 to 5.

Images