CN110947438A - Three-dimensional magnetic field parallel mixing microfluidic device and using method thereof - Google Patents
Three-dimensional magnetic field parallel mixing microfluidic device and using method thereof Download PDFInfo
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- CN110947438A CN110947438A CN201911286523.6A CN201911286523A CN110947438A CN 110947438 A CN110947438 A CN 110947438A CN 201911286523 A CN201911286523 A CN 201911286523A CN 110947438 A CN110947438 A CN 110947438A
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/502707—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by the manufacture of the container or its components
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/12—Specific details about manufacturing devices
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
- G01N35/00029—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor provided with flat sample substrates, e.g. slides
- G01N2035/00099—Characterised by type of test elements
- G01N2035/00158—Elements containing microarrays, i.e. "biochip"
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
- G01N2035/00465—Separating and mixing arrangements
- G01N2035/00534—Mixing by a special element, e.g. stirrer
Abstract
The invention discloses a three-dimensional magnetic field parallel mixing microfluidic device and a using method thereof, and belongs to the technical field of microfluidic chips. The device comprises two corresponding rectangular flat plates which are arranged in parallel, and a connecting structure at the tail end of each long flat plate, wherein the connecting structure is positioned at one end or two ends of each long flat plate; the long flat plates are respectively provided with a plurality of magnets; the magnets are uniformly arranged in an "S" shape on each long flat plate, and the magnets are staggered from each other in a vertical position. The device has the characteristics of portability, automation, low cost, low power consumption and the like, is convenient to use and flexible to apply, can meet the requirements of efficient mixing and reaction of samples with different fluxes, is easy to integrate other functional units, is favorable for further constructing a multi-sample real-time on-site analysis system, and has wide application prospect.
Description
Technical Field
The invention relates to a three-dimensional magnetic field parallel mixing microfluidic device and a using method thereof, belonging to the technical field of microfluidic chips.
Background
Magnetic beads are various in types, sizes and functions, and from nano-scale to micro-scale, the surfaces of the magnetic beads can be coated with various functional groups, antigens or antibodies, and the like, and the magnetic beads are widely applied to the treatment and analysis of various samples (such as cells, nucleic acids, proteins, and the like). For example, the immunomagnetic bead (IMB) technology is an immunological separation and detection technology based on specific antigen-antibody reaction. It uses magnetic beads coated by antibody as carrier, and makes the antibody combine with specific antigen in reaction liquor to form antigen-antibody compound, and said compound can be directionally moved under the action of external magnetic field so as to attain the goal of separating specific antigen. The method integrates the flexibility of a magnetic field control method and the high specificity of immunological reaction, and is widely applied to the aspects of cell separation, biomolecule purification, immunodetection and the like. In various applications involving magnetic beads, one of the key issues to be solved is the mixing efficiency of the magnetic beads with the sample and the processing throughput. The conventional magnetic bead mixing technology has high flux but low mixing efficiency, and finally influences the analysis and detection effect of a sample; or a more complex device is adopted to realize high-efficiency magnetic bead mixing (such as a gyroscope mixer and the like), but the device has the disadvantages of large volume, high cost, low flux and difficulty in integrating other operation units, thereby reducing the efficiency and the practicability of the method.
Disclosure of Invention
In order to solve the problems, the invention provides a three-dimensional magnetic field parallel mixing microfluidic control device and a using method thereof, and the device is particularly suitable for constructing a multi-sample parallel analysis microfluidic control system and can be applied to special environments such as multi-sample on-site rapid processing and microgravity.
The invention provides a three-dimensional magnetic field parallel mixing microfluidic device, which specifically comprises two corresponding long flat plates arranged in parallel and a long flat plate connecting structure at the tail end of each long flat plate, wherein the long flat plate connecting structure is positioned at one end or two ends of the short side of each long flat plate; the long flat plates are respectively provided with a plurality of magnets; the magnets are uniformly arranged in an "S" shape on each long flat plate, and the magnets are staggered from each other in a vertical position.
Specifically, in the three-dimensional magnetic field parallel hybrid microfluidic device, the magnets are embedded in the long flat plates, and are uniformly distributed on each long flat plate along the long side direction in an S shape and staggered with each other in the vertical position. Because the magnets are uniformly arranged on the upper long flat plate and the lower long flat plate in an S shape along the long side direction and are staggered with each other at the position vertical to the plate surface, the relative positions of each magnet and each mixing pool positioned between the two long flat plates are different and change regularly; in the process that the two long flat plates move back and forth along the long side direction, a three-dimensional magnetic field which changes circularly is generated in the chip mixing pool in the magnetic field area, and then magnetic beads are caused to reciprocate along the directions of an X axis, a Y axis and a Z axis in the magnetic field, so that the purpose of efficiently mixing the magnetic beads and a sample is achieved.
The three-dimensional magnetic field parallel hybrid microfluidic device further comprises a fixed support, and the fixed support is connected with the bottom end of the lower long flat plate.
The device comprises a fixed support, and is characterized by further comprising a driving motor matched with the fixed support, wherein the fixed support is driven by the driving motor to do one-dimensional reciprocating motion in the horizontal direction.
Specifically, the three-dimensional magnetic field parallel hybrid microfluidic device further comprises a clamp for fixing the microfluidic chip.
In addition, the invention also provides a using method of the three-dimensional magnetic field parallel mixing microfluidic device, which comprises the following steps:
① adding sample and magnetic beads into the mixing pool on the micro-fluidic chip;
② fixing the microfluidic chip to make each mixing pool located in the magnetic field area of the three-dimensional magnetic field parallel mixing microfluidic device;
③ the upper and lower long plates are driven by the driving motor to do one-dimensional reciprocating motion in the horizontal direction, so as to drive the magnets which are embedded on the long plates and are arranged in a staggered manner to do one-dimensional reciprocating motion in the horizontal direction, and further the region between the two long plates generates a periodic three-dimensional magnetic field, thereby driving the magnetic beads in the mixing tanks to do three-dimensional motion, and simultaneously realizing the parallel and high-efficiency mixing and reaction of the samples and the magnetic beads in each mixing tank.
The micro-fluidic chip of the device can be provided with a plurality of mixing pools, and different samples can be added into each mixing pool, so that the simultaneous mixing of a plurality of samples can be realized.
Compared with the prior art, the invention has the following beneficial effects:
1. the three-dimensional magnetic field parallel mixing microfluidic device adopts a microfluidic chip magnetic field control technology, and has the characteristics of portability, automation, low cost, low power consumption, multi-sample parallel processing, high-efficiency mixing and the like.
2. The three-dimensional magnetic field parallel mixing microfluidic control device designed by the invention is convenient to use and flexible to apply, can meet the requirements of efficient mixing and reaction of samples with different fluxes, and is easy to integrate other functional units so as to further construct a multi-sample real-time on-site analysis system.
Drawings
Fig. 1 is a schematic structural diagram of a three-dimensional magnetic field parallel hybrid microfluidic device.
Fig. 2 is a schematic structural diagram of a multi-mixing-cell microfluidic chip.
FIG. 3 is a schematic diagram of three-dimensional movement of magnetic beads in a mixing pool of a microfluidic chip.
FIG. 4 is a diagram showing the structure of a microfluidic chip including four mixing cells arranged in parallel.
FIG. 5 shows immunomagnetic beads and CD4 with different three-dimensional mixing time+Binding rate of T lymphocytes.
In the figure, 1, a magnet fixing unit; 11. an upper long flat plate; 12. an upper long flat plate magnet; 13. a lower long flat plate; 14. a lower long flat magnet; 15. a long flat plate connection structure; 2. fixing a bracket; 3. a drive motor; 4. and (4) a micro-fluidic chip.
Detailed Description
The following non-limiting examples will allow one of ordinary skill in the art to more fully understand the present invention, but are not intended to limit the invention in any way. In the schematic diagram of the device, the structural dimensions are not marked, the structural structure of the device is mainly embodied, and the structural proportion and the dimensions can be adjusted as required in the actual production and use process.
Example 1
The invention is further described below with reference to the accompanying drawings. Fig. 1 is a schematic structural diagram of a three-dimensional magnetic field parallel hybrid microfluidic device, and as can be seen from fig. 1, the device of the invention is composed of a magnet fixing unit 1, a fixing bracket 2, a driving motor 3 and a microfluidic chip 4.
Specifically, the magnet fixing unit 1 is composed of an upper long flat plate 11, an upper long flat plate magnet 12, a lower long flat plate 13, a lower long flat plate magnet 14 and a long flat plate connecting structure 15, wherein the upper long flat plate 11 and the lower long flat plate 13 are correspondingly arranged in parallel in the horizontal direction, and one end or two ends of the short sides of the upper long flat plate 11 and the lower long flat plate 13 are connected by the long flat plate connecting structure 15; the upper long flat plate 11 and the lower long flat plate 13 are respectively provided with a plurality of upper long flat plate magnets 12 and lower long flat plate magnets 14; the upper long flat plate magnets 12 and the lower long flat plate magnets 14 are uniformly arranged in an "S" shape on each long flat plate, and the upper long flat plate magnets 12 and the lower long flat plate magnets 14 are staggered from each other in vertical positions.
In the embodiment, the two rectangular flat plates have the length of 104mm, the width of 20mm and the thickness of 4 mm; the upper long flat magnet 12 and the lower field flat magnet 14 are both cylindrical neodymium iron boron permanent magnets with the same shape, the diameter of the magnets is 4mm, and the height of the magnets is 9 mm. The upper long flat plate magnet 12 and the lower field flat plate magnet 14 are respectively embedded in the upper long flat plate 11 and the lower long flat plate 13.
The bottom end of the magnet fixing unit 1 is connected with a driving motor 3 through a fixing support 2, and the fixing support 2 enables the long flat plate to do one-dimensional reciprocating motion in the horizontal direction under the driving of the driving motor 3. The driving motor consists of a miniature direct current motor and a 3V power supply.
Specifically, the three-dimensional magnetic field parallel hybrid microfluidic device further comprises a clamp for fixing the microfluidic chip.
FIG. 2 is a schematic diagram of the structure of a multi-mixing-cell microfluidic chip, which comprises a plurality of mixing cells (1, 2 … … n) arranged in parallel, and all the mixing cells are located in a magnetic field mixing area of the device. The mixing tank is of a closed structure, the material, shape, length and depth of the mixing tank are not limited, and at least one inlet and one outlet are arranged; the chip mixing pool loaded with the sample and the magnetic beads is placed in a magnetic field area between an upper long flat plate 11 and a lower long flat plate 13 of the device, after a driving motor 3 is started, the upper long flat plate 11 and the lower long flat plate 13 start one-dimensional reciprocating movement in the horizontal direction under the driving of a fixed support 2, and as magnets are uniformly distributed on the upper long flat plate and the lower long flat plate in an S shape along the long edge, the reciprocating movement of the long flat plates drives the magnets to generate periodic movement to generate a magnetic field which continuously changes on an X axis and a Y axis; meanwhile, because the magnets on the upper and lower long flat plates are staggered in the vertical position, the magnets generate a constantly changing magnetic field on the Z axis, so that the magnetic beads placed in the magnetic field region are three-dimensionally mixed with the sample in the circularly changing magnetic field, and the motion track is shown in fig. 3. When the two long flat plates do continuous reciprocating motion, the magnetic beads in the magnetic field mixing area and the sample are driven to be continuously subjected to three-dimensional mixing, and finally, sufficient contact and mixing are achieved.
Using the device of the present invention and a microfluidic chip containing four mixing cells arranged in parallel (see FIG. 4), immunomagnetic beads and human CD4 were examined for different mixing times+The combination rate of T lymphocytes, the mixing process is rapidly and automatically completed in four mixing tanks on the microfluidic chip; after mixing for 4 minutes, the binding rate of the immunomagnetic beads and the cells exceeds 60%, after mixing for 9 minutes, the binding rate reaches 67%, and the repeatability of a plurality of mixing pools is good, and the result is shown in fig. 5.
Claims (6)
1. A three-dimensional magnetic field parallel mixing microfluidic device is characterized in that: the device comprises two corresponding long flat plates which are arranged in parallel and a long flat plate connecting structure at the tail end of each long flat plate, wherein the long flat plate connecting structure is positioned at one end or two ends of the short side of each long flat plate; the long flat plates are respectively provided with a plurality of magnets; the magnets are uniformly arranged in an "S" shape on each long flat plate, and the magnets are staggered from each other in a vertical position.
2. The three-dimensional magnetic field parallel hybrid microfluidic device of claim 1, wherein: the magnets are embedded in the long flat plates, are uniformly distributed on each long flat plate along the long side direction in an S shape, and are staggered with each other in the vertical position.
3. The three-dimensional magnetic field parallel hybrid microfluidic device of claim 1, wherein: still include the fixed bolster, the fixed bolster is connected with long dull and stereotyped bottom down.
4. The three-dimensional magnetic field parallel hybrid microfluidic device of claim 3, wherein: the fixing support is driven by the driving motor to do one-dimensional reciprocating motion in the horizontal direction.
5. The three-dimensional magnetic field parallel hybrid microfluidic device of claim 1, wherein: the fixture is used for fixing the microfluidic chip.
6. Use of the device according to any of claims 1-5, characterized in that: the method comprises the following steps:
① adding sample and magnetic beads into the mixing pool on the micro-fluidic chip;
② holding the microfluidic chip such that each mixing cell is located in the magnetic field region of the device of claim 1;
③ the upper and lower long plates are driven by the driving motor to do horizontal one-dimensional reciprocating motion, so as to drive the magnets which are embedded on the long plates and are arranged in a staggered way to do horizontal one-dimensional reciprocating motion, and further the region between the two long plates generates a periodic three-dimensional magnetic field, thereby driving the magnetic beads in the mixing pools to do three-dimensional motion, and simultaneously realizing the paralleling, mixing and reaction of the samples and the magnetic beads in each mixing pool.
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CN106179544A (en) * | 2016-07-14 | 2016-12-07 | 大连海事大学 | Portable immunomagnetic beads three-dimensional hybrid device based on micro-fluidic chip and using method |
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CN106179544A (en) * | 2016-07-14 | 2016-12-07 | 大连海事大学 | Portable immunomagnetic beads three-dimensional hybrid device based on micro-fluidic chip and using method |
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