CN112007704A - Micro-fluidic chip and method for sorting micro-nano particles by inertial turbulence - Google Patents
Micro-fluidic chip and method for sorting micro-nano particles by inertial turbulence Download PDFInfo
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Abstract
The invention discloses a micro-fluidic chip and a method for sorting micro-nano particles by inertial turbulence, wherein the micro-fluidic chip comprises an upper substrate and a lower substrate; the upper layer substrate and the lower layer substrate are sealed and bonded together to form the microfluidic chip; the upper substrate is provided with a liquid inlet hole, an inlet liquid storage pool, an inertia flow channel, a sudden expansion flow channel, a turbulence obstacle, an inner outlet flow channel, an inner outlet liquid storage pool, an inner liquid outlet hole, an outer outlet flow channel, an outer outlet liquid storage pool and an outer liquid outlet hole; the liquid inlet hole is communicated with the inlet liquid storage tank, the inertia flow channel and the sudden expansion flow channel in sequence; then dividing the flow into two branches, wherein one branch is sequentially communicated with the inner outlet flow channel, the inner outlet liquid storage pool and the inner liquid outlet hole, and the other branch is sequentially communicated with the outer outlet flow channel, the outer outlet liquid storage pool and the outer liquid outlet hole; the turbulent flow obstacle is positioned in the sudden expansion flow channel. The micro-fluidic chip overcomes the technical bottleneck that the traditional inertial micro-fluidic chip is difficult to accurately sort micro-nano particles, and has the advantages of simple structure, good control precision and high flux.
Description
Technical Field
The invention relates to a micro-fluidic chip and a micro-nano particle sorting method based on inertial turbulence, belongs to the field of micro-fluidics, and can be used for precise control application of micro-nano biological particles and cells, such as precise capture, focusing, sorting and the like of micro-nano biological cells.
Background
The portable instant detection instrument has important application value in the aspects of dealing with on-site emergent acute disease diagnosis, malignant disease early screening and prognosis evaluation, promoting personalized medical development and the like, is an important carrier for the health requirements of people, and is highly valued by government departments of various countries in recent years. As a key technology of an instant detection instrument, the microfluidic chip has the advantages of high detection speed, high sensitivity, low cost, good integration and the like, is very suitable for the technical requirements of instant detection, and has become a research hotspot in the field at present.
The precise control (such as capture, focusing, separation and the like) of biological cells is an extremely important key step in the real-time detection pretreatment process, and the sensitivity and reliability of subsequent detection results are directly determined by the efficiency and the precision of sample processing. In view of this, the scholars at home and abroad have conducted a great deal of research and study on the cell manipulation method based on the microfluidic technology, and have reported a series of manipulation technologies based on physical fields such as electricity, magnetism, sound, light, etc. (i.e. active manipulation, such as dielectrophoresis, magnetophoresis, sound tweezers, light tweezers, etc.), manipulation technologies based on the self-structure of the microchannel (i.e. passive manipulation, such as deterministic lateral displacement, micro-barrier filtration, inertial microfluidic, etc.), and manipulation technologies based on active and passive mashup. The real-time controllability of active control is good, but the sample processing flux is low and the operation process is complex; the passive control has higher processing flux and does not need an external physical field, so the passive control has better integration advantage in a miniaturized device.
The inertial microfluidic technology utilizes the fluid inertia effect to induce cells to migrate in the flow channel under the action of inertia force so as to realize accurate control, has the advantages of simple flow channel structure, convenience in operation, high control precision and the like, and is widely concerned by domestic and foreign scholars. However, the fluid inertia effect has strong dependence on the apparent size of cells, and it is difficult to accurately control cells with high concentration and similar size (such as separating and capturing circulating tumor cells in blood), and the accurate acquisition of the cells has great application and scientific value for diagnosis, monitoring and treatment of some serious diseases.
Therefore, the traditional inertial microfluidic technology is broken through, the control performance of the micro-nano biological particles is improved, the biomedical application range of the inertial microfluidic is expanded, a research basis is provided for early screening and prognosis treatment of major diseases, and a technical support is provided for finally realizing industrial application of the inertial microfluidic chip.
Disclosure of Invention
The purpose of the invention is as follows: in order to overcome the defects in the prior art, the invention provides the micro-fluidic chip and the method for sorting the micro-nano particles by inertial turbulence, the chip has small volume, no need of sheath fluid, good control precision and high flux, and can meet the precise control application of the micro-nano biological particles and cells.
In order to achieve the purpose, the invention adopts the technical scheme that:
the invention provides a micro-fluidic chip for sorting micro-nano particles by inertial turbulence, which comprises an upper substrate and a lower substrate; the upper substrate and the lower substrate are sealed and bonded together to form the microfluidic chip.
The upper substrate is provided with a liquid inlet hole, an inlet liquid storage pool, an inertia flow channel, a sudden expansion flow channel, a turbulence obstacle, an inner outlet flow channel, an inner outlet liquid storage pool, an inner liquid outlet hole, an outer outlet flow channel, an outer outlet liquid storage pool and an outer liquid outlet hole; the liquid inlet hole, the inner liquid outlet hole and the outer liquid outlet hole are communicated with the outside and used for leading in and leading out the micro-nano particle solution; the liquid inlet hole is communicated with the inlet liquid storage tank, the inertia flow channel and the sudden expansion flow channel in sequence; then dividing the micro-nano particles into two branches, wherein one branch is sequentially communicated with the inner outlet flow channel, the inner outlet liquid storage pool and the inner liquid outlet hole, and the other branch is sequentially communicated with the outer outlet flow channel, the outer outlet liquid storage pool and the outer liquid outlet hole and used for accurately sorting the micro-nano particles; the turbulence obstacle is positioned in the sudden expansion flow channel.
The inertial microfluidic chip can be used for realizing inertial separation of more than two micro-nano particles with different sizes, and the size difference of the two micro-nano particles is not less than 3 mu m.
Preferably, the width of the flow channel of the inertia flow channel is greater than the height of the flow channel, so as to enhance the inertia focusing effect of the micro-nano particles in the inertia flow channel; the inertia flow channel is a bent flow channel and is used for generating a secondary flow effect perpendicular to the main flow direction of the cross section and enhancing the inertia migration effect of the micro-nano particles.
Preferably, the cross-sectional dimension of the inertial flow path is the same as the flow pathThe size relationship of the micro-nano particles is as follows:a/D h not less than 0.07, wherein a is the diameter of the micro-nano particles,D h is the height of the flow channel.
Preferably, the section of the inertia flow channel is rectangular, and the width-to-height ratio of the flow channel is 1.5-8.
Preferably, the curved flow passage has an Archimedes spiral structure.
Preferably, the ratio of the width of the sudden expansion flow channel to the width of the inertia flow channel is between 1.2 and 2.
Preferably, the turbulence barriers are one of rectangular or triangular, and the number is more than 1.
Preferably, the width ratio of the inner outlet flow channel to the outer outlet flow channel is 0.1-1.
Preferably, the material of the microfluidic chip is polydimethylsiloxane, thermoplastic polymer, glass or silicon chip.
The method for sorting the micro-nano particles by inertial turbulence comprises the following steps of:
(1) and injecting the micro-nano particle solution with different sizes into a liquid inlet hole of the micro-fluidic chip at a set flow rate by using a precise injection pump or an air pump, wherein the micro-nano particles are randomly dispersed in the flow channel when just entering the inertial flow channel.
(2) And the micro-nano particle solution in the inertia flow channel generates two secondary flow vortexes which are symmetrical up and down in the direction vertical to the main flow direction of the section. At the moment, the micro-nano particles are simultaneously subjected to the inertial lift force generated by the wall surface induction of the inertial flow channel and the secondary flow drag force generated by the turning of the solution in the spiral flow channel. The micro-nano particles gradually generate an inertial focusing effect under the influence of the inertial lift force and the secondary flow drag force and transversely move to different balance positions, and the specific expression is that the balance position of the large-size particles is slightly closer to the inner wall surface of the flow channel than the balance position of the small-size particles.
(3) When the micro-nano particles move through the sudden expansion flow channel and the turbulence obstacle, the large-size particles are quickly focused to the position close to the inner wall surface of the flow channel when bypassing the turbulence obstacle under the leading action of the inertial lift force, and the small-size particles are migrated to the outer wall surface when bypassing the turbulence obstacle under the leading action of the turbulence drag force, so that the balance position of the large-size particles and the small-size particles is increased.
(4) Finally, the large-size particles enter the inner outlet flow channel and flow out of the inner liquid outlet hole through the inner outlet liquid storage pool; and the small-size particles enter the outer outlet flow channel and flow out of the outer liquid outlet hole through the outer outlet liquid storage tank, so that the precise separation of the two different-size particles is realized.
The beneficial effects produced by the invention are as follows:
the micro-fluidic chip provided by the invention breaks through a general flow channel structure model of the traditional inertial micro-fluidic chip and designs a spiral flow channel chip with a novel flow channel structure. The sudden expansion flow channel and the turbulence obstacle structure are introduced into the spiral flow channel, so that the micro-nano particles not only receive the inertial lift effect generated by the wall surface of the flow channel and the secondary flow drag effect of the fluid perpendicular to the main flow direction of the cross section, but also receive the turbulence drag effect generated by the turbulence obstacle, the focusing balance position intervals of the micro-nano particles with different sizes are further increased, and the precise sorting of the micro-nano particles with different sizes is realized. Through the structural dimension parameters of changing the spiral flow channel, the sudden expansion flow channel and the turbulent flow obstacle, the inertia lift force, the secondary flow drag force and the turbulent flow drag force of the micro-nano particles can be changed, so that the micro-nano particles generate different balance positions, the sorting control precision and range of the micro-nano particles are enhanced, and the sorting device is beneficial to the accurate sorting of the micro-nano particles with more than three different sizes.
The chip disclosed by the invention is small in size, free of sheath fluid, good in control precision, high in flux, simple and convenient to manufacture, and has potential application value in the aspects of accurate control such as focusing, capturing and sorting of micro-nano particles (such as biological cells).
Drawings
Fig. 1 is a schematic 3D structure of a microfluidic chip;
FIG. 2 is a schematic view of the structure of an inertial flow path;
FIG. 3 is a schematic diagram of a micro-nano biological particle sorting experiment platform;
FIG. 4 is a schematic diagram of the random distribution of micro-nano biological particles on the section A-A of the spiral flow channel inlet;
FIG. 5 is a schematic view of inertial focusing of micro-nano biological particles on a section B-B of a spiral flow channel outlet;
fig. 6 is a schematic diagram of the migration and sorting of micro-nano biological particles in a sudden expansion channel.
Wherein, 1 is an upper layer substrate, 2 is a lower layer substrate, 3 is a liquid inlet hole, 4 is an inlet liquid storage tank, 5 is an inertia flow channel, 6 is a sudden expansion flow channel, 7 is a turbulence obstacle, 8 is an inner outlet flow channel, 9 is an inner outlet liquid storage tank, 10 is an inner liquid outlet hole, 11 is an outer outlet flow channel, 12 is an outer outlet liquid storage tank, and 13 is an outer liquid outlet hole.
Detailed Description
The preferred embodiments of the present invention will be described in conjunction with the accompanying drawings, and it will be understood that they are described herein for the purpose of illustration and explanation and not limitation.
As shown in fig. 1 and 2, the microfluidic chip provided by the invention is composed of an upper substrate 1 and a lower substrate 2; the upper substrate 1 and the lower substrate 2 are hermetically bonded together to form a microfluidic chip; the upper substrate 1 is provided with a liquid inlet hole 3, an inlet liquid storage pool 4, an inertia flow channel 5, a sudden expansion flow channel 6, a turbulence obstacle 7, an inner outlet flow channel 8, an inner outlet liquid storage pool 9, an inner liquid outlet hole 10, an outer outlet flow channel 11, an outer outlet liquid storage pool 12 and an outer liquid outlet hole 13; the liquid inlet hole 3, the inner liquid outlet hole 10 and the outer liquid outlet hole 13 are communicated with the outside and used for leading in and leading out the micro-nano particle solution; the liquid inlet hole 3 is communicated with an inlet liquid storage tank 4, an inertia flow channel 5 and a sudden expansion flow channel 6 in sequence; then, the liquid is divided into two branches, one branch is sequentially communicated with an inner outlet flow channel 8, an inner outlet liquid storage tank 9 and an inner liquid outlet hole 10, and the other branch is sequentially communicated with an outer outlet flow channel 11, an outer outlet liquid storage tank 12 and an outer liquid outlet hole 13; the method is used for precisely sorting micro-nano particles; the turbulence barrier 7 is located in the sudden expansion channel 6.
The inertia flow channel 5 is of an Archimedes spiral line structure, the inner diameter of the inlet of the flow channel is 10mm, and the outer diameter of the outlet of the flow channel is 30 mm. The cross section of the flow channel is rectangular, the width and the height are respectively 300 mu m and 50 mu m, and the width-height ratio of the flow channel is 6. Two kinds of target micro-nano biological particles to be sorted are polystyrene particles with diameters of 5 mu m and 15 mu m respectivelyAnd m is selected. The ratio of the diameter of the two biological particles to the height of the inertial flow channel 5 is respectively 0.1 and 0.3, which satisfies the requirementa/D h The particle focusing requirement is more than or equal to 0.07 (a is the diameter of the micro-nano particles,D h is the runner height). The width of the sudden expansion flow channel 6 is 500 μm, and the ratio of the width of the sudden expansion flow channel to the width of the inertia flow channel 5 is 1.67. The turbulence barriers are rectangular, 1 in number, and 300 μm and 400 μm in width and height, respectively. The width of the inner outlet flow channel 8 is 150 μm, the width of the outer outlet flow channel 11 is 350 μm, and the ratio of the widths of the inner outlet flow channel 8 and the outer outlet flow channel 11 is 0.43.
In the embodiment, the upper substrate 1 of the microfluidic chip is manufactured by using a standard soft lithography technology, the material is polydimethylsiloxane, the lower substrate 2 is a glass cover glass, and the upper substrate 1 and the lower substrate 2 are irreversibly bonded by an oxygen plasma cleaning process.
As shown in fig. 3, the microfluidic chip in this example was used to sort two polystyrene particles of 5 μm and 15 μm diameter. Firstly, a mixed particle solution is injected into the microfluidic chip by using a precision injection pump, and the flow rate is set to be 450 mu L/min. The mixed particle solution enters the inertia flow channel 5 from the inlet liquid storage tank 4 through the liquid inlet hole 3, and is in a random distribution state at the section A-A of the inlet of the inertia flow channel 5 (figure 4). Because the inertial flow channel 5 is in an archimedes spiral shape, microfluid in the flow channel generates two secondary flow vortexes which flow oppositely in the direction perpendicular to the main flow direction, and therefore the mixed particles are subjected to the inertial lift force generated by the wall surface induction of the inertial flow channel and the secondary flow drag force generated by the turning of the solution in the spiral flow channel in the flow channel. Then, the mixed particles are lifted in the inertia forceF L And secondary fluid drag forceF D Under the influence, the inertial focusing effect is gradually generated and the particles are transversely moved to different equilibrium positions, which is specifically shown in the way that the equilibrium position of the large-size particles is slightly closer to the inner wall surface of the flow channel than the equilibrium position of the small-size particles (figure 5), but the distance between the equilibrium positions of the two particles is smaller at the moment, so that the two particles cannot be accurately sorted. When the particles move through the sudden expansion flow channel 6 and the turbulence obstacle 7, the large-size particles are quickly focused to the near inner wall of the flow channel when bypassing the turbulence obstacle 7 under the leading action of the inertial lift forceAt the face, the small-sized particles migrate toward the outer wall surface while bypassing the turbulent obstacle 7 by the turbulent drag force, so that the equilibrium position of the large-sized particles and the small-sized particles is enlarged (fig. 6). Finally, the large-size particles enter the inner outlet flow channel and flow out of the inner liquid outlet hole through the inner outlet liquid storage pool; and the small-size particles enter the outer outlet flow channel and flow out of the outer liquid outlet hole through the outer outlet liquid storage tank, so that the precise separation of the two different-size particles is realized.
In conclusion, the inertial microfluidic chip provided by the embodiment has accurate inertial control performance of micro-nano particles, does not need sheath liquid supply, is small in size, simple and convenient to operate, and high in flux, can be used for controlling and applying efficient capture, focusing, sorting and the like of micro-nano biological cells, and has wide application value in aspects of integrating a microfluidic chip laboratory, a portable instant detection instrument and the like.
The above description is only of the preferred embodiments of the present invention, and it should be noted that: it will be apparent to those skilled in the art that various modifications and adaptations can be made without departing from the principles of the invention and these are intended to be within the scope of the invention.
Claims (10)
1. The utility model provides a micro-nano particle's micro-fluidic chip is selected separately to inertia vortex which characterized in that: comprises an upper substrate (1) and a lower substrate (2); the upper substrate (1) and the lower substrate (2) are hermetically bonded together to form a micro-fluidic chip;
the upper substrate (1) is provided with a liquid inlet hole (3), an inlet liquid storage pool (4), an inertia flow channel (5), a sudden expansion flow channel (6), a turbulence obstacle (7), an inner outlet flow channel (8), an inner outlet liquid storage pool (9), an inner liquid outlet hole (10), an outer outlet flow channel (11), an outer outlet liquid storage pool (12) and an outer liquid outlet hole (13);
the liquid inlet hole (3), the inner liquid outlet hole (10) and the outer liquid outlet hole (13) are communicated with the outside and used for leading in and leading out the micro-nano particle solution; the liquid inlet hole (3) is communicated with an inlet liquid storage pool (4), an inertia flow channel (5) and a sudden expansion flow channel (6) in sequence; then the flow is divided into two branches, one branch is sequentially communicated with an inner outlet flow channel (8), an inner outlet liquid storage pool (9) and an inner liquid outlet hole (10), and the other branch is sequentially communicated with an outer outlet flow channel (11), an outer outlet liquid storage pool (12) and an outer liquid outlet hole (13); the method is used for precisely sorting micro-nano particles; the turbulence obstacle (7) is positioned in the sudden expansion flow channel (6).
2. The microfluidic chip for sorting micro-nano particles by inertial turbulence according to claim 1, wherein: the width of the inertial flow channel (5) is greater than the height of the flow channel, and the inertial focusing effect of the micro-nano particles in the inertial flow channel (5) is enhanced; the inertia flow channel (5) is a bent flow channel and is used for generating a secondary flow effect perpendicular to the main flow direction of the cross section and enhancing the inertia migration effect of the micro-nano particles.
3. The microfluidic chip for sorting micro-nano particles by inertial turbulence according to claim 1, wherein: the cross section size of the inertia flow channel (5) and the size relation of the micro-nano particles in the flow channel are as follows:a/D h not less than 0.07, wherein a is the diameter of the micro-nano particles,D h is the height of the flow channel.
4. The microfluidic chip for sorting micro-nano particles by inertial turbulence according to claim 1, wherein: the section of the inertia flow channel (5) is rectangular, and the width-to-height ratio of the flow channel is 1.5-8.
5. The microfluidic chip for sorting micro-nano particles by inertial turbulence according to claim 2, wherein: the curved flow passage is of an Archimedes spiral line structure.
6. The microfluidic chip for sorting micro-nano particles by inertial turbulence according to claim 1, wherein: the ratio of the width of the sudden expansion flow channel (6) to the width of the inertia flow channel (5) is 1.2-2.
7. The microfluidic chip for sorting micro-nano particles by inertial turbulence according to claim 1, wherein: the turbulence barriers (7) are rectangular or triangular, and the number of the turbulence barriers is more than 1.
8. The microfluidic chip for sorting micro-nano particles by inertial turbulence according to claim 1, wherein: the width ratio of the inner outlet flow channel (8) to the outer outlet flow channel (11) is 0.1-1.
9. The microfluidic chip for sorting micro-nano particles by inertial turbulence according to claim 1, wherein: the material of the micro-fluidic chip is polydimethylsiloxane, thermoplastic polymer, glass or silicon chip.
10. A method for sorting micro-nano particles by inertial turbulence, which is characterized in that the microfluidic chip of claim 1 is used, and the method comprises the following steps:
(1) micro-nano particle solutions with different sizes are injected into a liquid inlet hole (3) of the micro-fluidic chip at a set flow rate by using a precise injection pump or an air pump, and the micro-nano particles are randomly dispersed in a flow channel when entering an inertia flow channel (5);
(2) the micro-nano particle solution in the inertial flow channel (5) generates two secondary flow vortexes which are vertically symmetrical in the direction perpendicular to the main flow direction of the section; at the moment, the micro-nano particles are simultaneously subjected to the inertial lift force generated by the wall surface induction of the inertial flow channel (5) and the secondary flow drag force generated by the turning of the solution in the spiral flow channel, the micro-nano particles gradually generate an inertial focusing effect under the influence of the inertial lift force and the secondary flow drag force and transversely move to different balance positions, and the specific expression is that the balance position of large-size particles is slightly closer to the inner wall surface of the flow channel than the balance position of small-size particles;
(3) when the micro-nano particles move through the sudden expansion flow channel (6) and the turbulence obstacle (7), the large-size particles are quickly focused to the position close to the inner wall surface of the flow channel when bypassing the turbulence obstacle (7) under the leading action of the inertial lift force, and the small-size particles are migrated to the outer wall surface when bypassing the turbulence obstacle (7) under the leading action of the turbulence drag force, so that the balance positions of the large-size particles and the small-size particles are increased;
(4) finally, the large-size particles enter the inner outlet flow channel (8) and flow out of the inner liquid outlet hole (10) through the inner outlet liquid storage pool (9); the small-size particles enter the outer outlet flow channel (11) and flow out of the outer liquid outlet hole (13) through the outer outlet liquid storage pool (12), and accurate separation of the two different-size particles is achieved.
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Cited By (4)
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| CN114100706A (en) * | 2021-10-18 | 2022-03-01 | 吉林大学 | Particle sorting method and system based on particle drift |
| CN114534809A (en) * | 2022-02-25 | 2022-05-27 | 河海大学常州校区 | Microfluidic particle control device with adjustable cross section shape and particle control method |
| CN114643088A (en) * | 2022-03-14 | 2022-06-21 | 常熟理工学院 | Micro-droplet generation chip based on Karman vortex street |
| CN115301303A (en) * | 2022-09-15 | 2022-11-08 | 中国矿业大学 | Multi-component mine dust separation micro-fluidic chip and classification concentration detection method thereof |
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| CN114643088A (en) * | 2022-03-14 | 2022-06-21 | 常熟理工学院 | Micro-droplet generation chip based on Karman vortex street |
| CN114643088B (en) * | 2022-03-14 | 2024-04-19 | 常熟理工学院 | Micro-droplet generation chip based on karman vortex street |
| CN115301303A (en) * | 2022-09-15 | 2022-11-08 | 中国矿业大学 | Multi-component mine dust separation micro-fluidic chip and classification concentration detection method thereof |
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