CN117816259A - A microfluidic chip structure and particle manipulation method based on swirl double stationary points - Google Patents
A microfluidic chip structure and particle manipulation method based on swirl double stationary points Download PDFInfo
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Abstract
The invention discloses a micro-fluidic chip structure based on rotational flow double standing points and a particle control method, and relates to the fields of micro-fluid, particle control methods and the like. The structure consists of three flow channels and a circular guide wall positioned in the middle area, and the particle control method is to uniformly spray fluid through the three flow channels, so that the microfluid forms a rotational flow low-pressure area on the left side and the right side of the circular guide wall, thereby sucking and capturing particles and stably rotating. The present structure and method provide for non-destructive manipulation of particles.
Description
Technical Field
The invention belongs to the field of microfluidics, relates to a particle control technology, and in particular relates to a microfluidic chip structure based on a rotational flow double standing point and a particle control method.
Background
Microparticle manipulation is a key technology in micromanipulation, and is also a core technology of a microfluidic chip, and is widely applied to the fields of biomedicine, material science and the like (Wu Chunhui, cheng Xin. Application of a microfluidic chip in single cell capture [ J ]. Science and technology guide, 2018,36 (16): 7.). Yang Chaoyong to Xiamen university et al proposes a microfluidic chip for precise manipulation and pairing of individual particles. The chip comprises a channel layer and a control layer, wherein the channel layer comprises a plurality of units for capturing and transferring single particles, and each unit comprises a capturing runner, a capturing chamber, a capturing gap, a transferring runner, a pairing chamber and a pairing gap. The control layer is positioned below the capturing flow channel and the pairing flow channel, is perpendicular to the capturing flow channel and the pairing flow channel and is isolated by a diaphragm. The microfluidic chip can be used for efficiently and accurately controlling the capturing and transferring of single particles, and after the single particles with different numbers are captured and transferred, the high-flux and high-efficiency single particle pairing can be realized, and the number and the types of paired particles are controllable. (Yang Chaoyong, liu Weizhi, li Xingrui, etc. A microfluidic chip for precisely manipulating and pairing single particles and application thereof, CN109722385A [ P ].2019 ]) but the method can only capture and transfer single particles, and cannot conduct multi-particle interaction research. Xing Xiaoxing et al discloses a microfluidic chip for cell-microbead capturing pairing, comprising a dielectrophoresis capturing part and a micro-well collecting part splitting buried layer electrode part. The dielectrophoresis force generated by electrifying the double-layer structure electrode of the dielectrophoresis capturing part and the periodic capturing groove of the electrode are utilized to capture the microbeads and the cells, so that double efficient capturing and pairing of the cells and the microbeads are realized, after capturing, dielectrophoresis excitation is stopped, the microbeads and the cells are settled into the micro-wells of the micro-well collecting part through the gravity effect, and the microbeads and the cells form one-to-one paired collection in the micro-wells of the micro-well collecting part. After the pairing is completed, a splitting electrode consisting of a double-layer structure electrode of the dielectrophoresis capturing part and a liquid electrode of the splitting buried layer electrode part is electrified to split cells. The invention employs an active control mechanism of dielectrophoresis to accomplish trajectory control and final capture of cells and microbeads, which enables handling of high throughput samples and shortens capture time, shortens development and manufacturing time and reduces costs (Xing Xiaoxing, liu Zhuzhu, cai Yao, etc. a microfluidic chip for cell-microbead capture pairing, CN115007231a [ P, 2022 ]. Cheng Xin and the like provide a cell array capturing and pairing microfluidic chip, which comprises cell pair units arranged in an array, wherein each cell pair unit is internally provided with a large micro-well, three small micro-wells, three groups of electrode pairs and shielding electrodes, the three small micro-wells are positioned in the large micro-wells and are arranged at sinking intervals, the shielding electrodes are positioned between adjacent small micro-wells, and the small micro-wells are positioned between the corresponding electrode pairs. The cell capturing and pairing microfluidic chip can capture three cells on the chip in sequence, so that a large-scale triple cell array is realized; three cells are taken out of the array, and the three cells are arranged in a pairing mode, so that great convenience and possibility are provided for researching cell-cell interactions such as paracellular secretion or cell fusion (Cheng Xin, tao Chaoran, wu Chunhui, and the like; a cell capturing and pairing microfluidic chip, CN114891628A [ P ] 2022. The method can realize the capture and control of two particles, but involves the use of an electric field, the device is complex, and the influence of the electric field on active biological cells is not clear. The method adopts a rotational flow control stagnation point method without an external force field, realizes the capture and control of double particles based on fluid dynamics and structural design, and has lower cost and higher safety.
The particle control method based on the rotational flow standing points does not cause mechanical damage to particles, has the advantages of simple control system, high sample processing flux and the like, but the research of controlling particles by using the rotational flow standing points in the rotational flow area is not mature, so that the further application of the method is limited. In micromanipulation using fluidic spots, safavieh et al, university of mcgill designed a multi-microtube, mobile, channel-free microfluidic device that achieved moving spots by varying the position and flow of the microtubes (Safavieh, m., et al, two-Aperture Microfluidic Probes as Flow Dipoles: theory and Applications (vol 5,11943,2015), scientific Reports, 2015.5). Yaxiaaer yanikun et al, university of osaka, japan, formed a reflow on a vertical plane by ejecting a fluid to the top of a microfluidic chip, and utilized the reflow to effect a spinning operation of different sized particles on the vertical plane (yanikun, y., kanda, and k. Morishima, hydrodynamic vertical rotation method for a single cell in an open space, microfluidics and Nanofluidics,2016.20 (5)). The swirl driving structure is proposed by the university of south China university Zhang Qin (Zhang Q, fan J, aoyama H.management of particles based on swirl [ J ]. Japanese Journal of Applied Physics,2018,57 (1): 017202 ]) and the like, two opposite placed microtubes generate swirl in the plane of the two microtubes by opposite jet flow, and the area near the center of the swirl has the characteristics of low pressure, low flow velocity and large pressure difference with the periphery of a swirl field. The cyclone center area has entrainment effect on particles in the cyclone flow area, and can capture the particles to enter the cyclone center; if the parameters of the swirl field are matched with those of the particles, the particles will rotate along their own axis under the action of viscous force and will not leave the center of the swirl. Three microtubes are arranged in the drainage basin, and particles in the swirling flow field can move along any direction in the drainage basin along with the swirling flow center while rotating by controlling the speed difference of the three microtubes. The capture, movement and rotation of particles can be realized by controlling the standing point of the rotational flow center.
The existing particle control method is difficult to simultaneously realize a series of operations such as particle capturing, moving and rotating, and a microfluidic chip based on a rotational flow standing point principle can only control single particles. Compared with the prior art, the scheme has the characteristics of simple control, strong anti-interference capability and easy integration. The research result is to realize a series of actions of particle posture adjustment, particle capture, movement and posture adjustment. On the basis, the invention provides a micro-fluidic chip structure based on a rotational flow double standing point and a particle control method, and two rotational flow low-pressure areas formed in a drainage basin are utilized to simultaneously capture and control two particles, so that the applicability of the micro-fluidic chip structure is enlarged.
Disclosure of Invention
Based on the condition that the nondestructive operation technology in the prior particle control technology is limited and the research on the rotational flow stagnation point method for controlling particles is insufficient for system maturation, the invention provides a micro-fluidic chip structure based on rotational flow double stagnation points and a particle control method, and designs a micro-fluidic chip structure capable of forming two rotational flow low-pressure region stagnation points and capturing and controlling two particles simultaneously. The double-dwell-point model can be used for fluid directional assembly of multiple particles in a solution, and provides a particle control method for multi-particle operation and assembly and basic research of particle-particle interaction.
The invention is realized at least by one of the following technical schemes.
The utility model provides a micro-fluidic chip structure based on two standing points of whirl, includes the micro-fluidic chip, the micro-fluidic chip includes the base plate, the base plate upper surface is equipped with three runner and guide wall, and second runner, third runner outlet are parallel to each other to be located the below of first runner outlet, the guide wall is located second runner, third runner central axis connection's center, and with the outlet terminal surface on a horizontal plane.
Further, the three flow channels are in the same plane, and the outlet end face of the first flow channel, the central axes of the second flow channel and the third flow channel form a closed rectangle.
Further, the cross section of the outlet ends of the three flow channels is rectangular, the interior of the three flow channels is changed into round holes through rectangular transition, and the flow of fluid is guaranteed to be equal in the flow channel structure transition process.
Further, the three flow channels are independently controlled by three syringe pumps, respectively, to provide microfluidics as fluid inlets.
Further, the guide wall is positioned between the second flow channel and the third flow channel, and the lower end of the guide wall is tangent to the connecting line of the outlet end surfaces of the second flow channel and the third flow channel.
Further, the guide wall is a circular guide wall.
Further, the inlets of the three flow channels are connected to a syringe.
Furthermore, positioning grooves are formed in two sides of the substrate, and the substrate is installed in the clamping platform through the positioning grooves.
Further, the microfluidic chip is mounted in a container by a leaf spring press.
The particle control method based on the rotational flow double-standing-point micro-fluidic chip structure is realized and is characterized by comprising the following steps of:
1) The microfluidic chip is fixed on the clamping platform by using the positioning grooves, and the flow input ports of the three flow channels (1, 3 and 4) are connected to the injector (12), and the injector (6) is connected with the hose and fixed in the clamping platform. Adjusting the micro-fluidic chip to ensure that the top surface of the chip is in a horizontal state;
2) Before a formal experiment, the air in the pipeline is emptied by utilizing the fluid so as to prevent bubbles from affecting the continuous input and output states of the fluid in the microtube;
3) Injecting microfluid into the three flow channels through the injector and the hose by adopting the injection pump, and forming two rotational flow areas in the flow field surrounded by the three flow channels; simultaneously, the liquid suction injector extracts liquid in the clamping platform;
4) Respectively throwing micron particles into the cyclone low-pressure area watershed;
5) Particles were observed to be trapped and stably maintained in the cyclone low pressure zone, when observed with a microscope;
6) The rotation speeds of the captured particles in the two flow areas are respectively controlled by changing the injection speeds of the fluid in the second flow passage and the third flow passage.
Further, the three flow channels are connected with the hoses through the adapter, the hoses are clamped in the groove channels of the clamping platform, and the three hoses are ensured to be on the same horizontal line as much as possible and are not mutually wound.
Further, the rotational flow strength at the standing point is controlled by adjusting the flow rates of the two flow channels at the lower side, so that the capture of particles with different sizes is realized.
Further, the structure is suitable for particle capturing and controlling with different sizes by changing the cross-sectional area of the flow channel, so that the applicable object of the structure is enlarged.
Compared with the prior art, the invention has the beneficial effects that:
1. the natural unloading of the fluid is considered, so that the interference of the speed outlet on the flow field is reduced as much as possible;
2. the cyclone double-resident-point structure can simultaneously capture and control two particles, so that the sample processing flux is improved.
Drawings
FIG. 1 is a schematic diagram of a microfluidic chip according to the present invention;
FIG. 2 is a cross-sectional view of the structure of a microfluidic chip according to the present invention;
FIG. 3 is a schematic diagram of a microfluidic chip experimental device based on a rotational flow double standing point;
FIG. 4 is an experimental microscope image of a swirling double stagnation particle manipulation;
wherein, 1-first runner, 3-second runner, 4-third runner, 2-guide wall, 5-base plate; the device comprises a 6-liquid suction injector, a 7-control panel, an 8-computer, a 9-clamping platform, a 10-microfluidic chip, an 11-microscope, a 12-liquid inlet injector and a 13-injection pump.
Detailed Description
In order that those skilled in the art will better understand the present invention, the following description will be given in detail with reference to the accompanying drawings and detailed description. It will be apparent that the described embodiments are only some, but not all, embodiments of the invention. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention. The present invention will be described in further detail with reference to examples and drawings, but embodiments of the present invention are not limited thereto.
As shown in fig. 1, the present embodiment provides a micro-fluidic chip structure based on a rotational flow double standing point, which comprises a micro-fluidic chip 10, wherein the micro-fluidic chip 10 comprises a substrate 5, three flow channels (1, 3, 4) and a guide wall 2 are arranged on the upper surface of the substrate 5, and the substrate 5, the three flow channels (1, 3, 4) and the guide wall 2 are in an integrated structure.
The outlets of the second flow channel 3 and the third flow channel 4 are parallel to each other and are positioned below the outlet of the first flow channel 1, and the outlet end face of the first flow channel 1 and the central axes of the second flow channel 3 and the third flow channel 4 form a closed rectangle; the guide wall 2 is positioned at the center of the central axis connection of the second flow channel 3 and the third flow channel 4 and is on the same horizontal plane with the outlet end face. The inlets of the three flow channels (1, 3, 4) connected to the injectors 12 are located at the side end faces of the base plate 5.
The cross-section of the outlet ends of the three flow channels (1, 3 and 4) is rectangular, and the interior of the flow channel is changed into a round hole from rectangular, so that the flow channel is convenient to adapt to an external round hose, and the flow of fluid is ensured to be equal in the flow channel structure conversion process.
As a preferred embodiment, threeThe cross-sectional dimensions of the outlet ends of the flow channels (1, 3, 4) are all 0.5 x 1.5mm 2 The size of the closed rectangle is 3X 6mm; the three flow channels (1, 3, 4) are independently controlled by three syringe pumps, respectively, to provide microfluidics as fluid inlets. Different cross-sectional areas of the flow channels can be designed according to the size of particles to be captured, so that the application range of the particle collector is enlarged. The guide wall 2 is a circular guide wall 2, and the radius is 0.5mm.
The microfluidic chip is of an open structure, an outlet adopts a natural unloading mode, and microfluid flows into the clamping disc container from the upper surface of the substrate 5. The upper surface of the base plate 5 is a horizontal plane, and deionized water on the upper surface of the base plate can overflow into the clamping platform container 9 after reaching a certain liquid level.
As a preferred embodiment, positioning grooves are formed on two sides of the base plate 5, and the base plate is installed in the clamping container through the positioning grooves. Besides clamping and fixing the microfluidic chip by using the positioning groove, the microfluidic chip is further fixed by using the plate spring pressing piece.
As shown in fig. 3, the microfluidic chip 10 is fixed on the clamping platform 9 by using a positioning groove, the inlets of three flow channels (1, 3, 4) of the microfluidic chip 10 are connected to the injector 12 by using an adapter external hose, the injection speed of fluid in the injector 12 is preset by using the control panel 7, the injector pump 13 is driven to feed and push the injector 12 to move at a certain speed, so that the liquid in the injector 12 enters the three flow channels at the set injection speed, and meanwhile, the liquid suction injector 6 can extract the liquid in the clamping platform 9, so that the flow stability of the fluid in the chip is ensured. Three injection pumps respectively provide liquid to three flow channels, and two cyclone low-pressure areas are formed on the left side and the right side of the guide wall 2, so that particles are captured. And a microscope 11 is arranged above the clamping platform 9 for real-time observation, and the microscope and the injection pump are connected to the computer 8, so that the condition of the swirling flow area on the microfluidic chip platform can be observed in real time.
Fig. 2 is a cross-sectional view of a micro-fluidic chip structure based on a rotational flow double standing point, and the flatness and smoothness of a middle rectangular area are required to be high during processing, so that the stability of a river basin in the middle rectangular area is required to be ensured during a particle manipulation experiment, and the rectangular size can be adjusted according to the size and the moving range of particles.
The micro-fluidic chip structure based on the rotational flow double standing points comprises the following steps:
1) The microfluidic chip (10) is fixed on the clamping platform (9) by using the positioning groove, and the three flow channels (1, 3, 4) are connected with the flow input ports of the injector (12), and the injector (6) is fixed in the clamping platform (9) through hose connection. Adjusting the micro-fluidic chip to ensure that the top surface of the chip is in a horizontal state;
2) Before a formal experiment, the air in the pipeline is emptied by utilizing the fluid so as to prevent bubbles from affecting the continuous input and output states of the fluid in the microtube;
3) Injecting microfluid into the three flow channels (1, 3, 4) at a certain speed (25-60 mm/s) through the injector (12) and the hose by adopting the injection pump (13), and forming 2 rotational flow areas in the flow areas surrounded by the three flow channels (1, 3, 4); simultaneously, the liquid suction injector (6) extracts the liquid in the clamping platform (9);
4) Respectively throwing micron particles into the cyclone low-pressure area watershed;
5) Particles were observed to be trapped and stably maintained in the cyclone low pressure zone, where they could be observed with the aid of a microscope;
6) By varying the injection velocity of the fluid in the two flow channels (3, 4) at the lower side, the rotational velocity of the captured particles in the two flow fields is controlled separately. The rotational flow strength at the standing point is controlled by adjusting the flow velocity of the two flow channels (3, 4) at the lower side, so that the particle capturing of different sizes or different qualities is realized.
Figure 4 shows a microscope image of a particle manipulation experiment based on a cyclone double standing point, the fluid speed of three channels is given, particles can be captured in a cyclone low-pressure area and can last for a plurality of hours without external interference, the liquid level is required to be noted in the experiment, and the particle used in the experiment is 100 mu m in diameter.
In the method, particle capturing with different sizes or different qualities can be realized by adjusting the flow velocity of the two flow channels at the lower side and controlling the rotational flow intensity at the standing point. Changing the feed rate of the syringe pump can change the fluid velocity in the flow channel and can change the capture position of the particles. The position of the circular guide wall can slightly move up, down, left and right so as to adjust the specific position of the particles captured.
The invention can be used for fluid directional assembly of multiple particles in solution, the rotational flow low-pressure areas on two sides can realize simultaneous control of cells and drug particles, and the distance between two standing points can be adjusted by changing the flow input of three flow channels or the position of a flow guide wall, so that the research on the interaction effect among the particles can be realized, and a new method is provided for the chemical analysis of the drugs.
The above description is only of the preferred embodiments of the present invention, but the protection scope of the present invention is not limited thereto, and any person skilled in the art can make equivalent substitutions or modifications according to the technical scheme and the inventive concept thereof within the scope of the present invention disclosed in the present invention, and all those skilled in the art belong to the protection scope of the present invention.
Claims (10)
1. The utility model provides a micro-fluidic chip structure based on two standing points of whirl, its characterized in that includes micro-fluidic chip (10), micro-fluidic chip (10) including base plate (5), base plate (5) upper surface is equipped with three runner (1, 3, 4) and water conservancy diversion wall (2), and the mutual parallel of second runner (3), third runner (4) export to be located the below of first runner (1) export, water conservancy diversion wall (2) are located the center of second runner (3), third runner (4) central axis connection, and with export terminal surface on a horizontal plane.
2. The micro-fluidic chip structure based on the rotational flow double standing points according to claim 1, wherein the three flow channels (1, 3, 4) are in the same plane, and the outlet end face of the first flow channel (1) forms a closed rectangle with the central axes of the second flow channel (3) and the third flow channel (4).
3. The micro-fluidic chip structure based on the rotational flow double standing points according to claim 1, wherein the cross section of the outlet ends of the three flow channels (1, 3 and 4) is rectangular, the interior is changed into a round hole from the rectangular, and the equal flow of the fluid is ensured in the process of changing the flow channel structure.
4. The micro-fluidic chip structure based on the rotational flow double standing points according to claim 1, wherein three flow channels (1, 3, 4) are independently controlled by three injection pumps, respectively, to provide micro-fluid as fluid inlets.
5. The micro-fluidic chip structure based on the rotational flow double standing points according to claim 1, wherein the guide wall (2) is positioned between the second flow channel (3) and the third flow channel (4), and the lower end of the guide wall (2) is tangent to a connecting line of the outlet end faces of the second flow channel (3) and the third flow channel (4).
6. The micro-fluidic chip structure based on the rotational flow double standing points according to claim 1, wherein the flow guiding wall (2) is a circular flow guiding wall (2).
7. A microfluidic chip structure based on a double standing point for swirling flow according to claim 1, characterized in that the flow input ports of the three flow channels (1, 3, 4) are connected to an injector (12).
8. The micro-fluidic chip structure based on the rotational flow double standing points according to any one of claims 1 to 7, wherein positioning grooves are arranged on two sides of the substrate (5), and the micro-fluidic chip structure is arranged in the clamping platform (9) through the positioning grooves.
9. The micro-fluidic chip structure based on rotational flow double standing points according to any of claims 1 to 7, characterized in that the micro-fluidic chip (10) is mounted in a container by a leaf spring press.
10. The method for controlling particles based on the rotational flow double-standing-point micro-fluidic chip structure according to claim 8 or 9 is realized, and is characterized by comprising the following steps:
1) The microfluidic chip (10) is fixed on the clamping platform (9) by utilizing a positioning groove, the flow input ports of the three flow channels (1, 3 and 4) are connected to the injector (12), and the injector (6) is connected with a hose and is fixed in the clamping platform; adjusting the micro-fluidic chip to ensure that the top surface of the chip is in a horizontal state;
2) Before a formal experiment, evacuating air in a pipeline by using fluid;
3) Injecting microfluid into the three flow channels through the injector (12) and the hose by adopting the injection pump (13), and forming two rotational flow areas in a flow field surrounded by the three flow channels; simultaneously, the liquid suction injector (6) extracts the liquid in the clamping platform (9);
4) Respectively throwing micron particles into the cyclone low-pressure area watershed;
5) Particles were observed to be trapped and stably maintained in the cyclone low pressure zone, when observed with a microscope;
6) The rotation speeds of the captured particles in the two flow areas are respectively controlled by changing the injection speeds of the fluid in the second flow passage (3) and the third flow passage (4).
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| US20060005634A1 (en) * | 2003-08-29 | 2006-01-12 | Schroeder Charles M | System and method for confining an object to a region of fluid flow having a stagnation point |
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| CN114836314A (en) * | 2022-06-01 | 2022-08-02 | 大连理工大学 | High-throughput microfluidic system for researching mechanical and biochemical signal induced single cell kinetic response and using method thereof |
| US20220347687A1 (en) * | 2019-06-13 | 2022-11-03 | Savelife Biotechnology Co. Limited | Separation method and apparatus for microvesicles |
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| US20060005634A1 (en) * | 2003-08-29 | 2006-01-12 | Schroeder Charles M | System and method for confining an object to a region of fluid flow having a stagnation point |
| AU2012245075A1 (en) * | 2011-04-20 | 2013-11-28 | 4Dx Limited | Method and device for trapping and analysing cells and the like |
| CN106622436A (en) * | 2016-12-26 | 2017-05-10 | 华南师范大学 | Light stream vortex array based material distributed control platform and control method |
| US20220347687A1 (en) * | 2019-06-13 | 2022-11-03 | Savelife Biotechnology Co. Limited | Separation method and apparatus for microvesicles |
| CN114836314A (en) * | 2022-06-01 | 2022-08-02 | 大连理工大学 | High-throughput microfluidic system for researching mechanical and biochemical signal induced single cell kinetic response and using method thereof |
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