CN110947437A - Portable microfluidic device for multi-sample and multi-step three-dimensional mixing and using method thereof - Google Patents
Portable microfluidic device for multi-sample and multi-step three-dimensional mixing and using method thereof Download PDFInfo
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- CN110947437A CN110947437A CN201911286517.0A CN201911286517A CN110947437A CN 110947437 A CN110947437 A CN 110947437A CN 201911286517 A CN201911286517 A CN 201911286517A CN 110947437 A CN110947437 A CN 110947437A
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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"
Abstract
The invention discloses a portable microfluidic device for multi-sample and multi-step three-dimensional mixing and a using method thereof, belonging to the technical field of magnetic field control microfluidic. The device comprises a three-dimensional magnetic field parallel mixing module and a magnetic bead horizontal moving module, wherein the three-dimensional magnetic field parallel mixing module comprises two corresponding rectangular flat plates which are arranged in parallel, a plurality of magnets are respectively arranged on the long flat plates, the magnets are uniformly distributed on each long flat plate in an S shape, and the magnets are staggered with each other in the vertical position; the magnetic bead horizontal moving module is composed of a strip-shaped magnet and a microfluidic chip position control assembly. The device can realize multistep three-dimensional mixing and reaction of a plurality of samples, is automatic, quick and efficient in the whole process, does not need professional operation, has the advantages of portability, low cost, easiness in further integration and the like, improves the practicability of a sample parallel processing method, and can be used for further constructing a multi-sample real-time field detection system.
Description
Technical Field
The invention belongs to the technical field of magnetic field control micro-fluidic, and particularly relates to a portable micro-fluidic device for multi-sample and multi-step three-dimensional mixing and a using method thereof.
Background
As a novel sample processing and detecting platform, the microfluidic technology has the characteristics of miniaturization, integration, automation and the like of an analysis device, and is a research hotspot in the technical field of sample analysis. The micro-processing technology is adopted to construct a micro-flow path system composed of a micro-scale liquid storage tank, a reaction tank, a channel and other structural units on a centimeter-scale chip, and then one or more reactions can be carried out on the micro-flow path system, so that the aims of rapidly processing and detecting samples are fulfilled. At the present stage, in order to ensure that the detection result is accurate and reliable, the conventional detection process can carry out parallel repeated detection on the same sample to be detected or different batches of samples to be detected under the same condition; in addition, the traditional detection method usually involves a multi-step mixed reaction process, and often requires professional staff to carry out the process step by step according to operation rules, and these practical requirements lead to large detection workload and poor detection result accuracy under the conditions of a large number of samples to be detected and complicated detection steps, so that the urgent requirements of real-time field detection are difficult to meet. Magnetic field control microfluidic technology can realize more complicated sample treatment and detection processes, but at present, it is difficult to provide an efficient multistep magnetic field control method, so that the final analysis and detection result is influenced, and meanwhile, the sample treatment flux is limited, so that the efficiency and the practicability of the method are reduced.
Disclosure of Invention
In order to solve the problems, the invention provides a portable microfluidic device for multi-sample and multi-step three-dimensional mixing and a using method thereof, and the device is particularly suitable for constructing a multi-sample real-time on-site detection system.
In order to achieve the purpose, the invention provides a portable micro-fluidic device for multi-sample and multi-step three-dimensional mixing, which comprises a three-dimensional magnetic field parallel mixing and magnetic bead horizontal moving module and a micro-fluidic chip position control assembly, and the automatic multi-sample and multi-step three-dimensional mixing based on a micro-fluidic chip is completed. Wherein:
the three-dimensional magnetic field parallel mixing and magnetic bead horizontal moving module 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, and a plurality of magnets are respectively arranged on the long flat plates; the magnets are uniformly arranged in an S shape on each long flat plate, and the magnets are staggered with each other in a vertical position; the magnets are distributed at one ends of the upper long flat plate and the lower long flat plate, and the other ends of the upper long flat plate and the lower long flat plate are magnetic bead horizontal moving modules; the long flat plate connecting structure is positioned on the short side of one end of the long flat plate, and the short side of the other end of the long flat plate is provided with a micro-fluidic chip position control assembly.
Furthermore, the magnets are embedded in the long flat plates, are uniformly arranged on each long flat plate along the long side direction in an S shape, and are staggered with each other in the vertical position.
Further, the magnetic bead horizontal movement module is a strip-shaped magnet, and the strip-shaped magnet is arranged on the upper long flat plate or the lower long flat plate.
Furthermore, the strip-shaped magnet is embedded in the upper long flat plate or the lower long flat plate, and the strip-shaped magnet is arranged along the long side direction of the long flat plate.
Furthermore, the device also comprises a long flat plate fixing support, and the long flat plate fixing support is connected with the bottom end of the lower long flat plate.
Further, the portable microfluidic device for multi-sample and multi-step three-dimensional mixing further comprises a long flat plate driving motor matched with the long flat plate fixing support, and the long flat plate fixing support is driven by the long flat plate driving motor to do one-dimensional reciprocating motion in the horizontal direction.
Furthermore, in the portable microfluidic device for multi-sample and multi-step three-dimensional mixing, the microfluidic chip position control assembly comprises a chip driving motor, a chip fixing clamp and a microfluidic chip, and the chip fixing clamp is driven by the chip driving motor to move horizontally; the chip fixing clamp drives the microfluidic chip to do a one-dimensional motion direction in the horizontal direction, and the motion direction of the microfluidic chip and the three-dimensional magnetic field parallel mixing module is vertical to each other in the horizontal position.
Another aspect of the invention is a method of use to protect the device described above, comprising the steps of:
① adding sample, magnetic beads and corresponding reagents into the mixing pool arranged in parallel on the microfluidic chip, and adding spacing reagents into the spacing pool;
② the first column of mixing cells (sample cells) of the microfluidic chip are all located in the three-dimensional magnetic field parallel mixing module region of the device;
③, the three-dimensional magnetic field parallel mixing module is started, the upper and lower long flat plates are driven by the motor to do one-dimensional reciprocating motion along the long side direction, so that the magnets embedded on the long flat plates and arranged in a staggered manner are driven to do one-dimensional reciprocating motion along the horizontal direction, a periodic three-dimensional magnetic field is generated in the area between the two long flat plates, the magnetic beads in the first row of mixing tanks are driven to do three-dimensional motion, and parallel and efficient mixing and reaction of the samples and the magnetic beads are realized;
④ after step ③, the microfluidic chip enters the magnetic bead horizontal movement module area, the long flat plate driving motor is turned off, the microfluidic chip position control assembly is turned on, and the chip driving motor drives the microfluidic chip to move along the horizontal direction perpendicular to the elongated magnet, so that the magnetic beads in the first row of mixing tanks enter the second row of mixing tanks through the isolation tanks, thereby realizing the separation of the magnetic beads from the solution in the first row of mixing tanks;
⑤ after step ④, turning off the chip driving motor, turning on the long flat plate driving motor, and horizontally moving the three-dimensional magnetic field parallel mixing module to the position of the second row of mixing tanks;
⑥ repeating steps ③ - ⑤ to make the magnetic beads pass through the subsequent mixing pool and isolation pool in sequence, and automatically realize the parallel mixing, separation and reaction of the magnetic beads and the solution in different mixing pools.
The invention has the following beneficial effects:
1. the invention takes the micro-fluidic chip and the magnetic field control device as the operation platform, adopts the three-dimensional magnetic field parallel mixing technology and combines the magnetic bead horizontal moving technology, can realize multi-step three-dimensional mixing and reaction of a plurality of samples, is automatic, quick and efficient in the whole process, does not need professional operation, and improves the practicability of the sample parallel processing method;
2. according to the invention, by adopting a method of a microfluidic technology and electromechanical integration equipment, the device has the advantages of portability, low cost, easiness in further integration and the like, and can be used for further constructing a multi-sample real-time field detection system.
Drawings
FIG. 1 is a schematic diagram of a portable microfluidic device configuration for multi-sample and multi-step three-dimensional mixing;
FIG. 2 is a schematic diagram of a microfluidic chip structure;
FIG. 3 is a top view of the apparatus of the present invention in two operating states;
FIG. 4 is a schematic diagram of a three-dimensional movement process of magnetic beads in a mixing pool of a microfluidic chip;
FIG. 5 is a block diagram of a microfluidic chip containing four sample analysis channels arranged in parallel;
FIG. 6 shows the results of nucleic acid extraction of bacterial lysates at different three-dimensional mixing times;
in the figure, 1, a three-dimensional magnetic field parallel mixing and magnetic bead horizontal moving module; 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; 16. a long flat plate fixing bracket; 17. a long flat plate drive motor; 2. a bar-shaped magnet; 3. a microfluidic chip position control assembly; 31. a microfluidic chip; 32. fixing a clamp on the chip; 33. the chip drives the motor.
Detailed Description
The following non-limiting examples are presented to enable those of ordinary skill in the art to more fully understand the present invention and 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 portable microfluidic device for multi-sample and multi-step three-dimensional mixing, as can be seen from fig. 1, the device of the invention is composed of a three-dimensional magnetic field parallel mixing and magnetic bead horizontal moving module 1, a strip-shaped magnet 2 and a microfluidic chip position control assembly 3. In addition, the device also comprises a micro-fluidic chip 31 in the using process.
Specifically, the three-dimensional magnetic field parallel mixing and magnetic bead horizontal moving module 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, a long flat plate connecting structure 15, a long flat plate fixing support 16 and a long flat plate driving motor 17. The upper long flat plate 11 and the lower long flat plate 13 are correspondingly arranged in parallel and are connected by a connecting structure 15 positioned at one end of the short side of the long flat plate; 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 evenly arranged in an "S" shape in the long side direction 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 the vertical position. In the embodiment, the two rectangular flat plates have the length of 104mm, the width of 20mm and the thickness of 4 mm; cylindrical neodymium iron boron permanent magnets with the same shape are distributed in the long flat plate 11 and the long flat plate 13, the diameter of the cylindrical neodymium iron boron permanent magnets is 4mm, and the height of the cylindrical neodymium iron boron permanent magnets is 9 mm. The long flat plate driving motor 17 is composed of a micro direct current motor and a 3V power supply, and the long flat plate is driven by the micro direct current motor of the long flat plate driving motor 17 through the long flat plate fixing support 16.
The magnetic bead horizontal movement module is a strip-shaped magnet 2, the strip-shaped magnet 2 is embedded in the upper long flat plate 11 or the lower long flat plate 13, and the strip-shaped magnet 2 is placed along the long side direction of the long flat plate.
Specifically, in a preferred scheme, the strip-shaped magnet 2 is a neodymium iron boron permanent magnet, the length is 30mm, the width is 10mm, and the thickness is 3 mm;
the portable microfluidic device for multi-sample and multi-step three-dimensional mixing further comprises a long flat plate fixing support 16, and the long flat plate fixing support 16 is connected with the bottom end of the lower long flat plate 13. The device also comprises a long flat plate driving motor 17 matched with the long flat plate fixing support 16, and the long flat plate fixing support 16 is driven by the long flat plate driving motor 17 to do one-dimensional reciprocating motion in the horizontal direction.
The microfluidic chip position control assembly 3 comprises a chip driving motor 33, a chip fixing clamp 32 and a microfluidic chip 31, wherein the chip fixing clamp 32 moves horizontally under the driving of the chip driving motor 33. The microfluidic chip position control assembly 3 is positioned on one side far away from the long flat plate connecting structure 15; the chip fixing clamp 32 drives the microfluidic chip 31 to make a one-dimensional motion direction in the horizontal direction, and the motion direction of the three-dimensional magnetic field parallel mixing and magnetic bead horizontal moving module 1 is mutually vertical in the horizontal position.
Example 2
FIG. 2 is a schematic diagram of a microfluidic chip according to the present invention, which includes a plurality of sample analysis channels (1, 2 … … n) arranged in parallel, each sample analysis channel including a plurality of mixing cells (1, 2 … … n), wherein isolation cells are disposed between the mixing cells; each sample analysis channel can complete a multi-step mixing reaction process of the same sample. The mixing tank and the isolation tank are of closed structures, the materials, the shapes, the lengths and the depths of the mixing tank and the isolation tank are not limited, and at least one liquid adding opening is formed. The device of the invention has two operating states (schematically shown in fig. 3), specifically as follows:
(1) and (3) parallel mixing of three-dimensional magnetic fields: placing a first row of mixing tanks (sample tanks) loaded with samples and magnetic beads between an upper long flat plate magnet 12 and a lower long flat plate magnet 14 of the device, setting a program, starting a long flat plate driving motor 17, and enabling an upper long flat plate 11 and a lower long flat plate 13 to start one-dimensional reciprocating motion under the drive of a long flat plate fixing support 16, wherein the magnets are uniformly distributed on the upper long flat plate and the lower long flat plate in an S shape along the long edge, and the reciprocating motion of the long flat plates drives the magnets to generate periodic one-dimensional reciprocating motion in the horizontal direction to generate magnetic fields which continuously change 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 trajectory is shown in fig. 4. 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.
(2) Horizontal movement of magnetic beads: after the long flat plate driving motor 17 moves the strip-shaped magnet 2 to the position of the first row of mixing tanks along the horizontal direction, the microfluidic chip position control assembly 3 is started, the chip driving motor 33 drives the microfluidic chip 31 to move along the direction vertical to the long edge of the strip-shaped magnet 2, and the magnetic beads in the first row of mixing tanks enter the second row of mixing tanks through the isolation tanks under the control of the strip-shaped magnet 2, so that the separation of the magnetic beads and the solution in the first row of mixing tanks is realized; the long flat plate drive motor 17 is then turned on to move the three-dimensional magnetic field control region to the location of the second row of mixing cells.
The device is continuously switched between two running states, and three-dimensional mixing and separation of magnetic beads in each row of mixing tanks are continuously completed until the experiment is finished, so that the multi-step efficient mixing and reaction process of a plurality of samples is realized.
Example 3
The microfluidic chip used in this example has a four-channel/five-liquid-cell structure, and a schematic diagram is shown in fig. 5. The device is adopted to extract nucleic acid from bacterial lysate under different three-dimensional mixing time, Vibrio harveyi lysate and magnetic silica beads are added into a first row of mixing tanks (sample tanks) of a microfluidic chip, a cleaning buffer solution 1, a cleaning buffer solution 2 and an elution buffer solution are respectively added into a second row to a fifth row of mixing tanks, and finally mineral oil is added into an isolation tank. Parameters are set according to an experimental flow, the results of nucleic acid extraction of the magnetic beads and the sample under different reaction times (first three-dimensional mixing) are inspected, the whole process is automatically completed at room temperature, and fig. 6 is a primary result of nucleic acid extraction concentration test under different three-dimensional mixing times, which shows that the nucleic acid extraction concentration is obviously increased along with the increase of the three-dimensional mixing time, and the repeatability among a plurality of analysis channels is better.
Claims (8)
1. A portable microfluidic device for multi-sample and multi-step three-dimensional mixing, characterized by: including three-dimensional magnetic field parallel mixing and magnetic bead horizontal migration module and micro-fluidic chip position control subassembly, wherein:
the three-dimensional magnetic field parallel mixing and magnetic bead horizontal moving module 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, and a plurality of magnets are respectively arranged on the long flat plates; the magnets are uniformly arranged in an S shape on each long flat plate, and the magnets are staggered with each other in a vertical position; the magnetic bodies are distributed at one ends of the upper long flat plate and the lower long flat plate, the other end of the upper long flat plate or the lower long flat plate is a magnetic bead horizontal moving module, the long flat plate connecting structure is located on the short side of one end of the long flat plate, and the short side of the other end of the long flat plate is provided with a micro-fluidic chip position control assembly.
2. The portable microfluidic device for multi-sample and multi-step three-dimensional mixing 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 portable microfluidic device for multi-sample and multi-step three-dimensional mixing of claim 1, wherein: the magnetic bead horizontal moving module is a strip-shaped magnet, and the strip-shaped magnet is arranged on the upper long flat plate or the lower long flat plate.
4. The portable microfluidic device for multi-sample and multi-step three-dimensional mixing of claim 3, wherein: the strip-shaped magnet is embedded in the upper long flat plate or the lower long flat plate, and the strip-shaped magnet is arranged along the long side direction of the long flat plate.
5. The portable microfluidic device for multi-sample and multi-step three-dimensional mixing of claim 1, wherein: the long flat plate fixing support is connected with the bottom end of the lower long flat plate.
6. The portable microfluidic device for multi-sample and multi-step three-dimensional mixing of claim 5, wherein: the device also comprises a long flat plate driving motor matched with the long flat plate fixing support, and the long flat plate fixing support is driven by the long flat plate driving motor to do one-dimensional reciprocating motion in the horizontal direction.
7. The portable microfluidic device for multi-sample and multi-step three-dimensional mixing of claim 1, wherein: the microfluidic chip position control assembly comprises a chip driving motor, a chip fixing clamp and a microfluidic chip, and the chip fixing clamp is driven by the chip driving motor to move horizontally; the chip fixing clamp drives the microfluidic chip to do a one-dimensional motion direction in the horizontal direction, and the motion direction of the microfluidic chip and the three-dimensional magnetic field parallel mixing module is vertical to each other in the horizontal position.
8. Use of a portable microfluidic device for multi-sample and multi-step three-dimensional mixing according to any of claims 1-7, wherein: the method comprises the following steps:
① adding sample, magnetic beads and corresponding reagents into the mixing pool arranged in parallel on the microfluidic chip, and adding spacing reagents into the spacing pool;
② locating the first array of mixing cells of the microfluidic chip in the three-dimensional magnetic field parallel mixing module region of the device of claim 1;
③, starting the three-dimensional magnetic field parallel mixing module, driving the upper and lower long flat plates to do one-dimensional reciprocating motion along the long side direction through the motor, thereby driving the magnets embedded on the long flat plates and arranged in a staggered manner to do one-dimensional reciprocating motion along the horizontal direction, further generating a periodic three-dimensional magnetic field in the area between the two long flat plates, driving the magnetic beads in the first row of mixing tanks to do three-dimensional motion, and simultaneously realizing the parallel mixing and reaction of the sample and the magnetic beads;
④ after step ③, the microfluidic chip enters the magnetic bead horizontal movement module area, the long flat plate driving motor is turned off, the microfluidic chip position control assembly is turned on, and the chip driving motor drives the microfluidic chip to move along the horizontal direction perpendicular to the elongated magnet, so that the magnetic beads in the first row of mixing tanks enter the second row of mixing tanks through the isolation tanks, thereby realizing the separation of the magnetic beads from the solution in the first row of mixing tanks;
⑤ after step ④, turning off the chip driving motor, turning on the long flat plate driving motor, and horizontally moving the three-dimensional magnetic field parallel mixing module to the position of the second row of mixing tanks;
⑥ repeating steps ③ - ⑤ to make the magnetic beads pass through the subsequent mixing pool and isolation pool in sequence, and automatically realize the parallel, mixing, separation and reaction of the magnetic beads and the solution in different mixing pools.
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| CN106248948A (en) * | 2016-07-14 | 2016-12-21 | 大连海事大学 | A kind of for the fluorescently-labeled portable micro fluidic device of Active immunity and using method thereof |
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| CN106248948A (en) * | 2016-07-14 | 2016-12-21 | 大连海事大学 | A kind of for the fluorescently-labeled portable micro fluidic device of Active immunity and using method thereof |
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