CN110387313B - Magnetophoretic microfluidic chip based on bubble mixing - Google Patents
Magnetophoretic microfluidic chip based on bubble mixing Download PDFInfo
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Abstract
The invention discloses a magnetophoretic microfluidic chip based on bubble mixing, which comprises a biological sample liquid inlet, a plurality of detection reagent liquid inlets, a mixed waste gas outlet, a filtered clean gas inlet, a process waste liquid outlet, a final product extracted biological sample outlet, a bubble breaking microstructure, an upper sealing rubber plug and a mixed working liquid cavity. According to the invention, through a bubble mixing mode, filtered clean gas is introduced to the bottom of the chip to generate bubbles in the reagent in the mixed working solution cavity, and the bubbles continuously rise in the reagent by utilizing the buoyancy of the bubbles to disturb surrounding fluid to generate micro-vortex, so that the effect of fully mixing the reagent and magnetic beads in the mixed working solution cavity is achieved. The invention solves the problem of uneven mixing of the biological sample liquid sample and the extraction magnetic beads in the micro-cavity, and is suitable for the field of microfluidic biological sample extraction and reagent mixing.
Description
Technical Field
The invention belongs to the technical field of micro-fluidic correlation, and relates to a micro-fluidic biological sample extraction and reagent mixing device.
Background
In gene analysis and disease detection, processes such as DNA extraction, amplification and isolation are often required, with DNA extraction being the most critical step. The traditional manual accounting extraction method is complex in operation, wastes a large amount of manpower, consumes a large amount of reagents, is difficult to popularize into large-scale repeated experiments because each step is not accurately controlled, and therefore an extraction process and a microfluidic chip are required to be combined to realize a micro total analysis system.
The micro total analysis system has the characteristics of realizing process integration, automation and miniaturization in a lab-on-a-chip, can greatly reduce the consumption of reagents, shortens the analysis time and improves the analysis and detection efficiency. Therefore, the method has gained wide attention in the fields of disease diagnosis, biochemical analysis and clinical experiments.
Disclosure of Invention
The invention provides a magnetophoresis microfluidic chip based on bubble mixing, and aims to solve the technical problems that an existing micro-extraction and biological sample extraction device is complex in operation, difficult to standardize and automate, needs a large amount of manpower and time to repeat experiments, and is small in equipment integration level, long in response time and high in price. According to the invention, through a bubble mixing mode, filtered clean gas is introduced to the bottom of the chip to generate bubbles in the reagent in the mixed working solution cavity, and the bubbles continuously rise in the reagent by utilizing the buoyancy of the bubbles to disturb surrounding fluid to generate micro-vortex, so that the effect of fully mixing the reagent and magnetic beads in the mixed working solution cavity is achieved.
The purpose of the invention is realized by the following technical scheme:
the utility model provides a magnetophoresis micro-fluidic chip based on bubble mixes, includes clean gas entry, process waste liquid export, final product extraction biological sample export, bubble destroy micro-structure, upper portion sealing rubber stopper and mixed work liquid chamber after biological sample inlet, multiple detect reagent inlet, mixed exhaust outlet, filtration, wherein:
the biological sample liquid inlet and the multiple detection reagent liquid inlets are connected with the upper part of the mixed working liquid cavity;
the upper end of the mixed working liquid cavity is provided with an upper sealing rubber plug, and a bubble breaking microstructure is arranged inside the mixed working liquid cavity;
the process waste liquid outlet and the final product extraction biological sample outlet are connected with the bottom of the mixed working liquid cavity;
the filtered clean gas inlet is connected with the middle position of the bottom of the mixed working liquid cavity, and the filtered clean gas is introduced into the solution from the bottom of the mixed working liquid cavity to form bubbles;
and the mixed waste gas outlet is connected with the top end of the mixed working liquid cavity to lead out the gas mixed in the chip.
The working principle is as follows:
the clean gas after filtration is introduced to the bottom of the chip, bubbles are generated in the reagent in the mixed working liquid cavity, and the bubbles rise by utilizing the buoyancy of the bubbles to disturb the surrounding fluid to generate micro-vortex under the condition of not introducing external interference, so that the effect of fully mixing the reagent in the mixed working liquid cavity is achieved. The bubbles are generated by introducing filtered clean gas into a filtered clean gas inlet which is connected with the bottom of the mixed working solution cavity and is positioned in the middle of the bottom. The solution passing through the inside of the mixed working liquid cavity rises by utilizing the buoyancy of bubbles to disturb surrounding fluid to generate micro-vortex, so that after the solution is fully mixed, the bubble breaking micro-structure formed by the left, middle and right inverted sharp triangular micro-structures inside the mixed liquid cavity actively eliminates the micro-structure, and the waste gas is discharged from a mixed waste gas outlet connected to the top end of the liquid cavity after being mixed in the chip. The invention can complete the complicated process of extracting the biological sample by the magnetic beads in the chip and has better mixing effect.
The buoyancy of the bubbles is utilized to rise to disturb the surrounding fluid to generate micro-vortex, so that the effect of full mixing is achieved.
Compared with the prior art, the invention has the following advantages:
1. the invention can complete the process of extracting the biological sample by the magnetic beads in the chip, has high mixing efficiency, does not need external equipment, and realizes the miniaturization, integration and automation of a trace extraction and biological sample extraction device.
2. The invention solves the problem of uneven mixing of the biological sample solution and the extraction magnetic beads in the micro-cavity, and is suitable for the field of microfluidic biological sample extraction and reagent mixing.
3. The magnetophoretic microfluidic chip disclosed by the invention has a simple working principle, bubbles are generated in a reagent in a mixed working liquid cavity by introducing filtered clean gas at the bottom of the chip, and the bubbles rise by utilizing the buoyancy of the bubbles to disturb surrounding fluid to generate micro-vortex under the condition of not introducing external interference, so that the effect of fully mixing the reagent in the mixed working liquid cavity is achieved.
4. The magnetophoretic microfluidic chip is formed by PMMA materials through machining by using a micro-milling machine, is easy to manufacture and machine, and is environment-friendly and harmless to human bodies.
5. The invention can complete the complicated process of extracting the biological sample by the magnetic beads in the magnetophoretic microfluidic chip, has good mixing effect and provides a basis for the automation of the next extraction technology.
6. The magnetophoretic microfluidic chip disclosed by the invention has the advantages of short reaction step standardization time, easiness in processing and low price, and meanwhile, the filtered clean gas is used for generating bubbles to achieve a mixing effect, and no other interference is introduced in the PCR reaction process.
7. The magnetophoretic microfluidic chip disclosed by the invention has the advantages that because the components are multi-layer combination with microstructures, different microstructures and micro-chambers can be designed for liquids with different densities and viscosities, and meanwhile, bubbles are generated by adopting gases with different pressures, so that the emulsification and mixing of various immiscible liquids can be realized, and the mixing effect is better.
Drawings
Fig. 1 is a schematic structural diagram of a magnetophoretic microfluidic chip based on bubble mixing according to the present invention.
Fig. 2 is a schematic three-dimensional structure diagram of a magnetophoretic microfluidic chip based on bubble mixing according to the present invention.
Detailed Description
The technical solution of the present invention is further described below with reference to the accompanying drawings, but not limited thereto, and any modification or equivalent replacement of the technical solution of the present invention without departing from the spirit and scope of the technical solution of the present invention shall be covered by the protection scope of the present invention.
The invention provides a magnetophoretic microfluidic chip based on bubble mixing, which is composed of a biological sample liquid inlet 1, a plurality of detection reagent liquid inlets, a mixed waste gas outlet 5, a filtered clean gas inlet 6, a process waste liquid outlet 7, a final product extraction biological sample outlet 8, a bubble breaking microstructure 9, an upper sealing rubber plug 10 and a mixed working liquid cavity 11, as shown in figures 1 and 2, wherein:
the multiple detection reagent liquid inlets are formed by sequentially arranging a No. 2 detection reagent liquid inlet 2, a No. 3 detection reagent liquid inlet 3 and a No. 4 detection reagent liquid inlet 4;
the mixed working liquid cavity 11 consists of a lower semicircular micro cavity and an upper rectangular micro cavity;
an upper sealing rubber plug 10 is arranged at the top of the rectangular micro-cavity at the upper part of the mixed working liquid cavity 11;
the bubble breaking microstructure 9 consists of three inverted sharp triangular microstructures, namely a left inverted sharp triangular microstructure, a middle inverted sharp triangular microstructure and a right inverted sharp triangular microstructure, and plays a role in actively breaking a large amount of bubbles to prevent the bubbles from overflowing;
the biological sample liquid inlet 1, the No. 2 detection reagent liquid inlet 2, the No. 3 detection reagent liquid inlet 3 and the No. 4 detection reagent liquid inlet 4 are connected with the rectangular micro-cavity at the upper part of the mixed working liquid cavity 11;
a bubble breaking microstructure 9 is arranged in the rectangular micro-cavity at the upper part of the mixed working liquid cavity 11;
the process waste liquid outlet 7 and the final product extraction biological sample outlet 8 are connected with the bottom center of the semicircular micro-cavity at the lower part of the mixed working liquid cavity 11;
the filtered clean gas inlet 6 is connected with the bottom center of the semicircular micro cavity at the lower part of the mixed working liquid cavity 11, and the filtered clean gas is introduced into the solution from the bottom of the semicircular micro cavity at the lower part of the mixed working liquid cavity 11 to form bubbles;
and the mixed waste gas outlet 5 is connected with the top end of the rectangular micro-cavity at the upper part of the mixed working liquid cavity 11 to lead out the mixed gas in the chip.
The specific working process of the magnetophoretic microfluidic chip based on bubble mixing is as follows:
and 8, injecting a certain amount of cleaning solution 1 into the mixed working solution cavity 11 from the No. 4 detection reagent liquid inlet 4, removing the magnet, resuspending the magnetic beads in the solution, inserting the magnetic beads into a steel needle through the filtered clean gas inlet 6, slowly opening a precision air pressure control gas source, and introducing the filtered clean gas to uniformly mix the solution in the liquid cavity, thereby completing the first cleaning step. After the cleaning process is finished, a numerically controlled mechanical arm is used for attaching a cylindrical strengthening magnet with the diameter of 10mm and the thickness of 5mm to the rear surface of the chip, the strengthening magnet moves in a scanning shape along the inside of the mixed working liquid cavity 11, magnetic beads with biological samples are adsorbed and fixed on the rear wall of the mixed working liquid cavity 11 through magnetic force, and the process needs 30s to complete sufficient adsorption. After the magnetic bead adsorption process is finished, inserting the steel needle into the process waste liquid outlet 7, and after the waste liquid after the reaction is completely discharged out of the chip, pulling out the steel needle from the process waste liquid outlet 7;
and 11, putting the chip into a centrifugal machine, enabling the final trace biological sample to be gathered at the lower end of the chip, adsorbing and fixing magnetic beads on the rear wall of the mixed working liquid cavity 11 through a strong magnet, and inserting a steel needle into the final product biological sample extraction outlet 8 after adsorption is finished to extract a certain amount of trace biological sample extraction liquid.
In the invention, the processing material of the magnetophoretic microfluidic chip is one or a combination of more of polydimethylsiloxane, polymethyl methacrylate, glass and high polymer resin.
In the invention, the bubble generating device adopts a small air pump to compress and filter clean gas instead of a pressure air bottle.
In the invention, the solution in the mixed solution cavity is not only the magnetic beads and the biological sample reagent, but also other chemical reagents, and is used for operations such as mixing, extraction and the like of other chemical reactions.
In the invention, the biological sample liquid inlet 1, the No. 2 detection reagent liquid inlet 2, the No. 3 detection reagent liquid inlet 3, the No. 4 detection reagent liquid inlet 4, the process waste liquid outlet 7 and the final product extraction biological sample outlet 8 are all covered by environment-friendly rubber sealing plugs, so that liquid can not leak.
Claims (5)
1. The utility model provides a magnetophoresis micro-fluidic chip based on bubble mixes, its characterized in that magnetophoresis micro-fluidic chip includes biological sample inlet, multiple detect reagent inlet, mixed exhaust outlet, the clean gas entry after filtering, process waste liquid export, final product extraction biological sample export, bubble destroy micro-structure, upper portion sealing rubber stopper and mixed work liquid cavity, wherein:
the biological sample liquid inlet and the multiple detection reagent liquid inlets are connected with the upper part of the mixed working liquid cavity;
the upper end of the mixed working solution cavity is provided with an upper sealing rubber plug, and a bubble breaking microstructure is arranged in the mixed working solution cavity;
the bubble breaking microstructure consists of three inverted sharp triangular microstructures on the left, the middle and the right;
the process waste liquid outlet and the final product extraction biological sample outlet are connected with the bottom of the mixed working liquid cavity;
the filtered clean gas inlet is connected with the right center of the bottom of the mixed working liquid cavity, and the filtered clean gas is introduced into the solution from the bottom of the mixed working liquid cavity to form bubbles;
and the mixed waste gas outlet is connected with the top end of the mixed working liquid cavity to lead out the gas mixed in the chip.
2. The magnetophoretic microfluidic chip based on bubble mixing of claim 1, wherein the plurality of detection reagent liquid inlets are formed by sequentially arranging a No. 2 detection reagent liquid inlet, a No. 3 detection reagent liquid inlet and a No. 4 detection reagent liquid inlet.
3. The bubble-mixing-based magnetophoretic microfluidic chip of claim 1 or 2, wherein the biological sample liquid inlet, the multiple detection reagent liquid inlets, the process waste liquid outlet, and the final product extraction biological sample outlet are all covered by an environmentally-friendly rubber sealing plug.
4. The bubble mixing-based magnetophoretic microfluidic chip of claim 1, wherein the mixed working fluid chamber is comprised of a lower semicircular microcavity and an upper rectangular microcavity.
5. The magnetophoretic microfluidic chip based on bubble mixing of claim 1, wherein the processing material of the magnetophoretic microfluidic chip is one or a combination of polydimethylsiloxane, polymethyl methacrylate, glass and polymer resin.
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| AU2015225844B2 (en) * | 2014-03-07 | 2019-10-24 | National Research Council Of Canada | Centrifugal microfluidic chip control |
| CN104984773B (en) * | 2015-05-25 | 2016-10-05 | 重庆大学 | A kind of method of strengthening fluid mixing in microchannel |
| CN105349401B (en) * | 2015-10-14 | 2018-03-27 | 安徽易康达光电科技有限公司 | A kind of multi-functional integrated micro-flow control foranalysis of nucleic acids chip and preparation and analysis method |
| CN105597846B (en) * | 2015-10-26 | 2017-11-28 | 深圳华迈兴微医疗科技有限公司 | The magnetic microparticle chemiluminescence micro-fluidic chip that a kind of D dimers quantitatively detect |
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