CN109837204B - Micro-fluidic chip detection system and method integrating cell sorting and focusing - Google Patents
Micro-fluidic chip detection system and method integrating cell sorting and focusing Download PDFInfo
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Abstract
The invention discloses a micro-fluidic chip detection system and a method for integrated cell sorting and focusing, comprising a viscoelastic fluid sample injection device, a sample injection device, a waste liquid collection device, a high-speed image collection device, a micro-fluidic chip, a sample collection device and a microcomputer, wherein the micro-fluidic chip is packaged by a flow channel layer and a basal layer, and the flow channel layer is provided with a spiral flow channel, a bifurcation flow channel, a lower branch flow channel, an upper branch flow channel, a viscoelastic fluid sample injection flow channel, a spiral mixing flow channel and a focusing main flow channel; the invention realizes the sorting of cells according to the dimensional characteristics, the mixing and the blending of the viscoelastic carrier liquid, and the focusing of the sorted rare cells at a single balance position in the center of the cross section of the flow channel; the accuracy of the counting detection is improved.
Description
Technical Field
The invention relates to a biological particle control and detection technology based on a micro-fluidic chip, in particular to a micro-flow control chip rare cell count detection system integrating a spiral flow passage inertial separation technology, a spiral flow passage mixing technology and a viscoelastic inertial direct flow passage focusing technology.
Background
Rare cells which are rare and have important clinical value exist in body fluids such as blood, urine, pleural effusion and the like, but counting and detection of the rare cells are very challenging due to the very low content of the rare cells. At present, the common rare cell detection technology in clinical laboratories mainly comprises histochemical staining, immunohistochemical and flow cytometry technologies and the like, and the technologies are relatively mature, but have the defects of incapability of reaching detection requirements in sensitivity, very complicated operation, high instrument price and the like.
The micro-fluidic technology can realize accurate control of micro-fluid, has the advantages of accurate control, high efficiency, low cost and the like, and is widely applied to aspects of mixing, focusing, sorting, detection and the like of cells. At present, the cell control mode based on micro-flow control is mainly divided into an active mode and a passive mode, and the active mode has the advantages of high universality, high accuracy and the like by means of external field acting forces such as sound, magnetism, electricity and the like, but the passive mode does not need the auxiliary action of the external field, has the advantages of high flux, low cost, easiness in operation and the like, and has a good application prospect in a micro-flow control chip. Many microfluidic chips have been developed today that employ passive manipulation, such as by sorting cells according to size characteristics and then counting the sorted cells directly. However, after the sorted cells enter a new flow channel, the original focusing state of the cells can be influenced, so that the detection precision is influenced; in addition, when the cell counting is detected, the cell is difficult to keep single-position focusing, the problems that the cells are side by side, a plurality of cells pass through the counting detection area at the same time and the like can occur, and the accuracy of the cell counting detection is reduced. Therefore, the realization of more accurate focusing and detection has very important significance in the field of cell sorting and detection.
Disclosure of Invention
The invention aims to: in order to overcome the defects in the prior art, the invention provides a micro-fluidic chip detection system and a method for integrated cell sorting focusing, wherein the system integrates a spiral flow passage inertial sorting mixing technology and a viscoelastic inertial focusing technology in a direct flow passage; the spiral inertial flow channel realizes the separation of cells according to the size characteristics thereof by means of inertial effect and Dean flow, and has high flux and rapid separation, and the high flow speed is beneficial to the mixing of the bearing liquid in the next step; the spiral mixing flow channel realizes the mixing and the allocation of the viscoelastic carrier liquid, so that the solution can be mixed under the Dean flow action in the positive and negative directions, the expected viscoelastic action is shown, and the follow-up direct flow channel is helped to stably focus the sorted rare cells at a single balance position in the center of the flow channel section; the accuracy of counting detection is improved, and the purpose of high-accuracy detection is achieved.
The technical scheme is as follows: in order to achieve the above purpose, the invention adopts the following technical scheme:
the utility model provides a micro-fluidic chip detecting system of integrated cell separation focus, includes viscoelastic fluid sampling device, sample sampling device, waste liquid collection device, image acquisition device, micro-fluidic chip, sample collection device and microcomputer, wherein:
the microfluidic chip comprises a runner layer and a substrate layer, wherein the runner layer and the substrate layer are packaged from top to bottom; the flow channel layer comprises a spiral flow channel, a branched flow channel, a lower branched flow channel, an upper branched flow channel, a viscoelastic fluid sample injection flow channel, a spiral mixing flow channel and a focusing main flow channel; one end of the spiral flow channel is provided with a sample inlet, and the other end of the spiral flow channel is connected with the bifurcation flow channel; the branch flow passage is provided with two outlets, one outlet is connected with the inlet of the lower branch flow passage, the other outlet is connected with the inlet of the upper branch flow passage, the lower branch flow passage is positioned at one side far away from the spiral flow passage, and the upper branch flow passage is positioned at one side close to the spiral flow passage; the outlet of the lower branch flow passage is a waste liquid outlet;
the flow channel rotating direction of the spiral flow channel is anticlockwise rotation or clockwise rotation; the spiral mixing flow passage comprises more than two mixing branch flow passages, wherein two adjacent mixing branch flow passages are connected through a sudden expansion structure flow passage, and the flow passage rotation directions between the two adjacent mixing branch flow passages are opposite; the focusing main flow channel is a straight flow channel;
the inlet of the viscoelastic fluid sample injection flow channel is a viscoelastic sample inlet, and the outlet of the upper branch flow channel and the outlet of the viscoelastic fluid sample injection flow channel are both connected to the inlet of the spiral mixing flow channel; the outlet of the spiral mixing flow channel is connected with the inlet of the focusing main flow channel; the outlet of the focusing main flow channel is a rare cell outlet;
the viscoelastic fluid sample injection device is connected with the viscoelastic sample inlet through a first micro tube; the sample injection device is connected with the sample inlet through a second micro tube; the waste liquid collecting device is connected with the waste liquid outlet through a third micro tube; the sample collection device is connected with a rare cell outlet through a fourth microtube; the image acquisition device is in communication connection with the microcomputer through a data line; the image acquisition device is positioned above the focusing main runner and is close to the rare cell outlet.
Preferably: the focusing main flow passage is an inertial straight flow passage with a square section.
Preferably: the area of the inlet section of the focusing main flow passage is larger than that of the outlet section of the spiral mixing flow passage.
Preferably: the viscoelastic solution injected into the viscoelastic fluid sample injection device is polyvinylpyrrolidone solution or hyaluronic acid solution.
Preferably: the number of the mixing branch flow passages is two, and the directions of the two mixing branch flow passages relative to the flow passage with the abrupt expansion structure are opposite.
The method comprises the steps that a sample injection device conveys diluted samples into a spiral flow channel through a first microtube, and meanwhile, a viscoelastic fluid injection device starts to inject viscoelastic solution; the cells are subjected to inertia effect and Dean flow in the spiral flow channel, rare cells and red blood cells are gathered at different balance positions, then the rare cells are conveyed into the upper branch flow channel through the bifurcation flow channel to obtain rare cell solution, the red blood cells are conveyed into the lower branch flow channel, and the red blood cells enter the waste liquid collecting device through the third microtube; the rare cell solution and the injected viscoelastic solution meet and enter the spiral mixing flow channel together, the rare cell solution and the viscoelastic solution are mixed in the spiral mixing flow channel under the action of Dean flow, and the uniformly mixed carrier liquid shows expected viscoelasticity; then when the bearing liquid enters the focusing main runner; in the focusing main runner, because the fluid is subjected to the combined action of the viscoelastic effect and the inertia effect of the mixed bearing liquid, rare cells are gradually collected to the center of the runner in the solution, so that focusing at a single balance position of the center of the runner is realized, then image recording is carried out on the rare cells collected in the runner through an image acquisition device, the recorded pictures are transmitted to a microcomputer, analysis processing is carried out on the rare cells through the microcomputer, and the solution after image acquisition enters a sample collection device.
Compared with the prior art, the invention has the following beneficial effects:
the fluid gathers rare cells and other cells at different balance positions in the spiral flow channel due to inertial effect and Dean flow effect, then the rare cells are led into the upper branch flow channel through the bifurcation flow channel, and the other cells are led into the lower branch flow channel; the rare cell solution and the high-viscosity elastic solution which are injected originally meet and enter the spiral mixing flow channel together, and the rare cell solution and the high-viscosity elastic solution are mixed in the spiral mixing flow channel under the action of Dean flow; when the mixed fluid enters the focusing main flow passage, the flow velocity of the fluid is reduced due to the enlarged cross-sectional area of the flow passage; in the focusing main flow channel, because the fluid is subjected to the combined action of the viscoelastic effect and the inertial effect, rare cells are gradually collected to the center of the flow channel in the solution, so that focusing at a single balance position in the center of the flow channel is realized, the possibility that the original focusing state of the cells is influenced after the rare cells enter a new flow channel is avoided, and the problems that the cells are side by side, a plurality of cells pass through a counting detection area at the same time and the like which can occur originally are solved; recording the aggregated rare cells in the flow channel by a high-speed image acquisition device, transmitting the recorded pictures to a microcomputer, and analyzing and processing the forms, the number, the surface characteristics and the like of the rare cells by the microcomputer; the method realizes the more accurate acquisition of counting detection results of the sorted rare cells.
Drawings
FIG. 1 is a schematic diagram of a microfluidic chip system capable of implementing cell sorting, focusing at a cell center and detection according to the present invention;
FIG. 2 is a schematic diagram of a micro flow channel chip according to the present invention;
FIG. 3 is a schematic diagram of the principle of inertial separation of a spiral flow channel according to the present invention;
FIG. 4 is a schematic diagram of the separation principle of the bifurcated flow channel of the present invention;
FIG. 5 is a schematic diagram of the spiral mixing flow channel principle of the present invention;
FIG. 6 is a schematic diagram of the structure of the spiral mixing channel and the focusing main channel of the present invention;
FIG. 7 is a schematic diagram of the principle of focusing cells in a viscoelastic inertial flow channel in a focusing main flow channel according to the present invention;
in the figure: 11. the device comprises a viscoelastic fluid sample injection device, 111, a first microtube, 12, a sample injection device, 121, a second microtube, 13, a waste liquid collecting device, 131, a third microtube, 14, a high-speed image collecting device, 15, a microfluidic chip, 151, a runner layer, 152, a basal layer, 16, a sample collecting device, 161, a fourth microtube, 17, a microcomputer, 171, a data line, 21, a spiral runner, 211, a sample inlet, 22, a bifurcation runner, 23, a lower bifurcation runner, 231, a waste liquid outlet, 24, an upper bifurcation runner, 25, a viscoelastic fluid sample injection runner, 251, a viscoelastic sample inlet, 26, a spiral mixing runner, 27, a focusing main runner, 271, a rare cell outlet, 28, a sudden expansion structure runner, 31, a spiral runner outer wall surface, 32, a spiral runner inner wall surface, 33, dean flow, 41, rare cells, 42, red cells, 51, a mixing runner outer wall surface, 52, a mixing runner inner wall surface, 53, a viscoelastic solution, 54, a rare cell solution.
Detailed Description
The present invention is further illustrated in the accompanying drawings and detailed description which are to be understood as being merely illustrative of the invention and not limiting of its scope, and various equivalent modifications to the invention will fall within the scope of the appended claims to the skilled person after reading the invention.
An integrated cell sorting focusing micro-fluidic chip detection system, as shown in fig. 1 and 2, comprises a viscoelastic fluid sample injection device 11, a sample injection device 12, a waste liquid collection device 13, an image acquisition device 14, a micro-fluidic chip 15, a sample collection device 16 and a microcomputer 17, wherein:
as shown in fig. 2, the microfluidic chip 15 is formed by packaging a runner layer 151 and a substrate layer 152 from top to bottom; the flow channel layer 151 includes a spiral flow channel 21, a branched flow channel 22, a lower branched flow channel 23, an upper branched flow channel 24, a viscoelastic fluid sample injection flow channel 25, a spiral mixing flow channel 26, and a focusing main flow channel 27; one end of the spiral flow channel 21 is provided with a sample inlet 211, and the other end is connected with the branch flow channel 22; the branched flow passage 22 is provided with two outlets, one of the outlets is connected with the inlet of the lower branched flow passage 23, the other outlet is connected with the inlet of the upper branched flow passage 24, the lower branched flow passage 23 is positioned at one side far away from the spiral flow passage 21, and the upper branched flow passage 24 is positioned at one side close to the spiral flow passage 21; the outlet of the lower branch flow channel 23 is a waste liquid outlet 231;
the flow passage rotation direction of the spiral flow passage 21 is counterclockwise rotation or clockwise rotation; the spiral mixing flow channel 26 includes more than two mixing branch flow channels, two adjacent mixing branch flow channels are connected through a sudden expansion structure flow channel 28, the flow channel rotation directions between the two adjacent mixing branch flow channels are opposite, the number of the mixing branch flow channels is two, and the directions of the two mixing branch flow channels about the sudden expansion structure flow channel 28 are opposite; the focusing main flow channel 27 is an inertial flow channel with a square cross section.
The inlet of the viscoelastic fluid sample injection flow channel 25 is a viscoelastic sample inlet 251, and the outlet of the upper branch flow channel 24 and the outlet of the viscoelastic fluid sample injection flow channel 25 are both connected to the inlet of the spiral mixing flow channel 26; the outlet of the spiral mixing flow passage 26 is connected with the inlet of the focusing main flow passage 27; the outlet of the focusing main flow channel 27 is a rare cell outlet 271; the area of the inlet cross section of the focusing main flow passage 27 is larger than the area of the outlet cross section of the spiral mixing flow passage 26.
The viscoelastic fluid sample injection device 11 is connected with the viscoelastic sample inlet 251 through the first microtube 111; the viscoelastic solution 53 injected into the viscoelastic fluid sample injection device 11 is a polyvinylpyrrolidone (PVP) solution or a Hyaluronic Acid (HA) solution. The sample introduction device 12 is connected with the sample inlet 211 through a second microtube 121; the waste liquid collecting device 13 is connected with the waste liquid outlet 231 through the third micro tube 131; the sample collection device 16 is connected to the rare cell outlet 271 via a fourth microtube 161; the image acquisition device 14 is in communication connection with the microcomputer 17 through a data line 171; the image acquisition device 14 is located above the focusing main flow channel 27, and the image acquisition device 14 is close to the rare cell outlet 271.
The viscoelastic fluid sample injection device 11 and the sample injection device 12 are injection pumps, the waste liquid collection device 13 and the sample collection device 16 are test tubes, and the image collection device 14 is a camera.
The main working procedure of the invention is as follows: the sample introduction device 12 conveys the diluted blood sample into the spiral flow channel 21 through the first microtube 111, and simultaneously the viscoelastic fluid introduction device 11 starts to inject the viscoelastic solution 53; the cells are subjected to inertial effect and Dean flow in the spiral flow channel 21, the rare cells 41 and the red blood cells 42 are gathered at different balance positions due to different volumes, then the rare cells 41 are conveyed into the upper branch flow channel 24 through the branch flow channel 22 to obtain a rare cell solution 54, the red blood cells 42 are conveyed into the lower branch flow channel 23, and the red blood cells 42 enter the waste liquid collecting device 13 through the third microtube 131; the rare cell solution 54 meets the originally injected viscoelastic solution 53 and enters the spiral mixing flow channel 26 together, the rare cell solution 54 and the viscoelastic solution 53 are mixed in the spiral mixing flow channel 26 under the action of the cross-section Dean flow, and the mixed particle carrier liquid shows the expected viscoelastic effect; then, when the mixed fluid enters the focusing main flow channel 27; in the focusing main flow channel 27, since the fluid is subjected to the combined action of the viscoelastic effect and the inertial effect, the rare cells 41 gradually collect in the solution to the center of the flow channel, so that focusing at a single balance position in the center of the flow channel is realized, then the collected rare cells in the flow channel are subjected to image recording by the image acquisition device 14, the recorded pictures are transferred to the microcomputer 17, the morphology, the number, the surface characteristics and the like of the rare cells are analyzed and processed by the microcomputer 17, and the solution after image acquisition enters the sample collection device 16.
As shown in fig. 3, which is a schematic diagram of the principle of inertial separation of the spiral flow channel according to the present invention, in the spiral flow channel 21, the movement of the fluid can be decomposed in the cross-sectional direction of the flow channel. In the cross-section direction of the flow channel, the cell receives shear induced inertial lift force directed to the wall surface and wall surface induced inertial lift force directed to the center of the flow channel, which are collectively referred to as inertial lift force F, the fluid in the center of the flow channel is faster than the flow velocity of the surrounding fluid, and the centrifugal force received is also greater, so that the fluid in the center of the flow channel flows to the outer wall surface 31 of the spiral flow channel, and the fluid near the outer wall surface 31 of the spiral flow channel flows back along the upper and lower wall surfaces, so that a Dean flow 33 is generated in the cross section of the flow channel, and the Dean flow 33 generates a drag force F on the cell D Acting; as in the cells at (1) (2) (3) (4) (5) (6) of FIG. 3, the forces to which each are subjected are different, only the inertial lift force F and the Dean flow drag force F are applied at (1) D The two forces are balanced by the mutual cancellation of the two forces, and the cells are balanced and focused. In addition, inertial lift force F and Dean flow drag force F D The size of (2) is related to the size of the cells, so the final sizeThe larger rare cells equilibrate closer to the inner wall surface 32 of the spiral flow path, while the red blood cells equilibrate closer to the outer wall surface 31 of the spiral flow path.
FIG. 4 is a schematic diagram showing the separation principle at the branching flow channel; at the end of the spiral flow channel 21, the rare cells 41 and the blood cells 42 have reached their respective equilibrium positions with each other due to the combined action of the above two forces; when the cells enter the branched flow channel 22, the wall surface induced inertial lift force which is applied to the cells and is directed to the center of the flow channel is reduced due to the widening of the flow channel, so that the cells approach the wall surface, and the distance between the rare cells 41 and the blood cells 42 is also increased; then the fluid enters the bifurcation, the rare cells 41 enter the upper branch flow passage, and the blood cells 42 enter the lower branch flow passage, thereby completing the cell sorting.
The sorted rare cell solution 54 and the viscoelastic solution 53 enter the mixing flow channel together; as shown in fig. 2, at the interface of the upper branch flow channel 24, the viscoelastic fluid injection flow channel 25 and the spiral mixing flow channel 26, the flow channel at the interface is opposite to the flow direction of the liquid entering the upper branch flow channel 24, so that the fluid entering the flow channel at the interface at the rare cell solution 54 sorted by the spiral flow channel 21 still has the inertial lift force F and Dean flow drag force F generated by the spiral flow channel 21 D After entering the spiral mixing flow channel 26, the spiral mixing flow channel 26 also generates corresponding inertial lift force F and Dean flow, and at this time, the inertial lift force F and Dean flow drag force F generated by the spiral flow channel 21 D And the inertial lift force F and Dean flow drag force generated by the spiral mixing flow channel 26, the interaction of these four forces causes the rare cell solution 54 and the high visco-elastic solution 53 to mix at the interface. In the mixing channel, as shown in fig. 5, the movement of the fluid can be decomposed in the cross-section direction of the channel, since the channel is a curved channel, the fluid in the center of the channel is faster than the surrounding fluid, and the centrifugal force is also greater, so that the highly viscoelastic solution 53 in the center of the channel flows toward the outer wall surface 51 of the mixing channel, and the rare cell solution 54 near the outer wall surface 51 of the mixing channel flows back along the upper and lower wall surfaces, thus generating Dean flow 33 in the cross section of the channel; thin filmThe cell solution 54 and the high-viscosity elastic solution 53 move transversely in the flow channel under the action of centrifugal force and Dean flow, so that the two solutions are mixed, then enter the spiral flow channel 28 with the opposite rotation direction through the flow spiral flow channel, the original directions of the centrifugal force and the Dean flow become the original opposite directions, the comprehensive mixing effect is improved, and finally the rare cell solution with viscoelasticity is obtained.
As shown in fig. 6, the end of the spiral mixing channel 26 enters the focusing main channel 27, the focusing main channel 27 is an inertial channel with a square cross section, and the inlet cross section of the focusing main channel 27 is larger than the cross section of the spiral mixing channel 26, so that the effect of increasing the cross section of the channel and reducing the flow rate is achieved. Meanwhile, the Dean flow of the spiral mixing flow channel 26 has a preliminary focusing effect on rare cells, so that the focusing speed of the rare cells in the follow-up focusing main flow channel 27 can be accelerated to a certain extent.
As shown in fig. 2, the spiral mixing channel 26 is connected with the focusing main channel 27 through a vertical pipeline, that is, the direction of the focusing main channel 27 is perpendicular to the tangential direction of the connection of the spiral mixing channel 26, so when the liquid flows from the spiral mixing channel 26 to the connection, the inertial lift force F and Dean flow drag force generated by the spiral mixing channel 26 on the liquid are turned, on one hand, the mixing of rare cells in the spiral mixing channel 26 is accelerated by the design, on the other hand, the inertial lift force F and Dean flow drag force generated by the spiral mixing channel 26 are reduced, so that the liquid enters the focusing main channel 27, and the influence of the inertial lift force F and Dean flow drag force generated by the spiral mixing channel 26 is reduced, thereby facilitating the focusing of the rare cells, as shown in fig. 7; the rare cells in the fluid are dragged along the flowing direction to accelerate to the same speed as the surrounding fluid, and meanwhile, the rare cells are sheared by the fluid in the direction perpendicular to the flowing direction to generate inertial lift force, so that the rare cells are focused at the balance position within a short distance; and because the rare cells are subjected to the viscoelastic effect, the rare cells are focused at a single balance position in the center of the flow channel.
And then recording the collected rare cells in the flow channel through a high-speed image acquisition device, transmitting the recorded pictures to a microcomputer, and analyzing and processing the collected picture information in the microcomputer by utilizing software to obtain the morphology, the number, the surface characteristics and the like of the rare cells. The method realizes the more accurate acquisition of counting detection results of the sorted rare cells.
The invention realizes the sorting of cells according to the dimensional characteristics, the mixing and the blending of the viscoelastic carrier liquid, and the focusing of the sorted rare cells at a single balance position in the center of the cross section of the flow channel; the accuracy of the counting detection is improved.
The foregoing is only a preferred embodiment of the invention, it being 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 present invention, and such modifications and adaptations are intended to be comprehended within the scope of the invention.
Claims (6)
1. A micro-fluidic chip detection system integrating cell sorting focusing is characterized in that: including viscoelastic fluid sampling device (11), sample sampling device (12), waste liquid collection device (13), image acquisition device (14), micro-fluidic chip (15), sample collection device (16) and microcomputer (17), wherein:
the microfluidic chip (15) comprises a runner layer (151) and a substrate layer (152), wherein the runner layer (151) and the substrate layer (152) are packaged from top to bottom; the flow channel layer (151) comprises a spiral flow channel (21), a branch flow channel (22), a lower branch flow channel (23), an upper branch flow channel (24), a viscoelastic fluid sample injection flow channel (25), a spiral mixing flow channel (26) and a focusing main flow channel (27); one end of the spiral flow channel (21) is provided with a sample inlet (211), and the other end of the spiral flow channel is connected with the bifurcation flow channel (22); the branched flow passage (22) is provided with two outlets, one outlet is connected with the inlet of the lower branched flow passage (23), the other outlet is connected with the inlet of the upper branched flow passage (24), the lower branched flow passage (23) is positioned at one side far away from the spiral flow passage (21), and the upper branched flow passage (24) is positioned at one side close to the spiral flow passage (21); the outlet of the lower branch flow passage (23) is a waste liquid outlet (231);
the flow passage rotating direction of the spiral flow passage (21) is anticlockwise rotation or clockwise rotation; the spiral mixing flow channel (26) comprises more than two mixing branch flow channels, two adjacent mixing branch flow channels are connected through a sudden expansion structure flow channel (28), and the flow channel rotation directions between the two adjacent mixing branch flow channels are opposite; the focusing main runner (27) is a direct current runner;
the inlet of the viscoelastic fluid sample injection runner (25) is a viscoelastic sample inlet (251), and the outlet of the upper branch runner (24) and the outlet of the viscoelastic fluid sample injection runner (25) are connected to the inlet of the spiral mixing runner (26); the outlet of the spiral mixing runner (26) is connected with the inlet of the focusing main runner (27); the outlet of the focusing main runner (27) is a rare cell outlet (271);
the viscoelastic fluid sample injection device (11) is connected with the viscoelastic sample inlet (251) through a first microtube (111); the sample injection device (12) is connected with the sample inlet (211) through a second microtube (121); the waste liquid collecting device (13) is connected with the waste liquid outlet (231) through a third micro tube (131); the sample collection device (16) is connected to a rare cell outlet (271) through a fourth microtube (161); the image acquisition device (14) is in communication connection with the microcomputer (17) through a data line (171), and the image acquisition device (14) is a camera; the image acquisition device (14) is located above the focusing main runner (27), and the image acquisition device (14) is close to the rare cell outlet (271).
2. The integrated cell sorting focused microfluidic chip detection system of claim 1, wherein: the focusing main flow channel (27) is an inertial straight flow channel with a square cross section.
3. The integrated cell sorting focused microfluidic chip detection system of claim 2, wherein: the area of the inlet section of the focusing main flow passage (27) is larger than that of the outlet section of the spiral mixing flow passage (26).
4. The integrated cell sorting focused microfluidic chip detection system of claim 3, wherein: the viscoelastic solution (53) in the viscoelastic fluid sample injection device (11) is a polyvinylpyrrolidone solution or a hyaluronic acid solution.
5. The integrated cell sorting focused microfluidic chip detection system of claim 4, wherein: the number of the mixing branch flow passages is two, and the directions of the two mixing branch flow passages are opposite relative to the flow passage (28) with the abrupt expansion structure.
6. A detection method using the integrated cell sorting focused microfluidic chip detection system of claim 1, the detection method not involving diagnosis and treatment of disease, characterized in that: the sample injection device (12) conveys the diluted sample into the spiral flow channel (21) through the second microtube (121), and meanwhile, the viscoelastic fluid injection device (11) starts to inject the viscoelastic solution (53); the cells are subjected to inertia effect and Dean flow in the spiral flow channel (21), rare cells (41) and red blood cells (42) are gathered at different balance positions, then the rare cells (41) are conveyed into the upper branch flow channel (24) through the branch flow channel (22) to obtain rare cell solution (54), the red blood cells (42) are conveyed into the lower branch flow channel (23), and the red blood cells enter the waste liquid collecting device (13) through the third microtube (131); the rare cell solution (54) and the injected viscoelastic solution (53) meet and enter the spiral mixing flow channel (26) together, and the rare cell solution (54) and the viscoelastic solution (53) are mixed in the spiral mixing flow channel (26) by generating transverse movement under the action of a cross-section Dean flow; when the mixed fluid enters a focusing main runner (27); in a focusing main runner (27), due to the combined action of the viscoelasticity effect and the inertia effect of fluid, rare cells (41) are gradually converged to the center of the runner in the solution, so that focusing at a single balance position of the center of the runner is realized, then image recording is carried out on the rare cells converged in the runner through an image acquisition device (14), recorded pictures are transferred to a microcomputer (17), analysis processing is carried out on the rare cells through the microcomputer (17), and the solution after image acquisition enters a sample collection device (16).
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| WO2022194020A1 (en) * | 2021-03-17 | 2022-09-22 | Mgi Tech Co., Ltd. | Sample sorting device and method |
| CN113952993B (en) * | 2021-11-23 | 2022-09-20 | 中北大学 | Multistage inertial microfluidic blood sample processing chip integrating micro mixer and Tesla valve |
| CN114073997B (en) * | 2021-11-30 | 2022-07-05 | 南京林业大学 | Microfluidic chip and method for realizing rapid and accurate cell sorting at low flow rate |
| CN114700126B (en) * | 2022-05-06 | 2024-03-19 | 季华实验室 | Microfluidic sorting chip |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB0615546D0 (en) * | 2006-08-03 | 2006-09-13 | Proimmune Ltd | MHC Oligomer, Components Therof, And Methods Of Making The Same |
| CN101563104A (en) * | 2006-02-01 | 2009-10-21 | 约翰霍普金斯大学 | Polypeptide-nucleic acid conjugate for immunoprophylaxis or immunotherapy for neoplastic or infectious disorders |
| CN103923825A (en) * | 2014-04-17 | 2014-07-16 | 东南大学 | Microfluidic chip system integrating cell sorting and detection |
| CN105462834A (en) * | 2016-01-22 | 2016-04-06 | 苏州汶颢芯片科技有限公司 | Tumor cell capturing micro-fluidic chip and tumor cell capturing method |
| EP3072595A1 (en) * | 2015-03-24 | 2016-09-28 | European Molecular Biology Laboratory | Microfluidic sorting device |
| CN108495809A (en) * | 2016-01-28 | 2018-09-04 | 明策生物医学科技私人有限公司 | Use the multisection type target cell concentration of microfluidic device |
| CN109456875A (en) * | 2018-11-02 | 2019-03-12 | 东南大学 | The rare cell multipass sort micro-fluidic device of integrated inertia and certainty lateral displacement technology |
| CN209816114U (en) * | 2019-04-12 | 2019-12-20 | 南京林业大学 | Micro-fluidic chip detection system integrating cell sorting and focusing |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100329986A1 (en) * | 2009-01-06 | 2010-12-30 | Greenbaum Linda E | Adult Hepatic Progenitor Cells and Methods of Use Thereof |
-
2019
- 2019-04-12 CN CN201910293917.8A patent/CN109837204B/en active Active
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101563104A (en) * | 2006-02-01 | 2009-10-21 | 约翰霍普金斯大学 | Polypeptide-nucleic acid conjugate for immunoprophylaxis or immunotherapy for neoplastic or infectious disorders |
| GB0615546D0 (en) * | 2006-08-03 | 2006-09-13 | Proimmune Ltd | MHC Oligomer, Components Therof, And Methods Of Making The Same |
| CN103923825A (en) * | 2014-04-17 | 2014-07-16 | 东南大学 | Microfluidic chip system integrating cell sorting and detection |
| EP3072595A1 (en) * | 2015-03-24 | 2016-09-28 | European Molecular Biology Laboratory | Microfluidic sorting device |
| CN105462834A (en) * | 2016-01-22 | 2016-04-06 | 苏州汶颢芯片科技有限公司 | Tumor cell capturing micro-fluidic chip and tumor cell capturing method |
| CN108495809A (en) * | 2016-01-28 | 2018-09-04 | 明策生物医学科技私人有限公司 | Use the multisection type target cell concentration of microfluidic device |
| CN109456875A (en) * | 2018-11-02 | 2019-03-12 | 东南大学 | The rare cell multipass sort micro-fluidic device of integrated inertia and certainty lateral displacement technology |
| CN209816114U (en) * | 2019-04-12 | 2019-12-20 | 南京林业大学 | Micro-fluidic chip detection system integrating cell sorting and focusing |
Non-Patent Citations (1)
| Title |
|---|
| 微流控器件中血细胞行为的研究与分离技术;陈园园;高扬;熊成章;李疆;陈皓生;;中国科学:技术科学(第07期);第684-696页 * |
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