CN209974734U - Micro-fluidic chip and system for separating various cells - Google Patents
Micro-fluidic chip and system for separating various cells Download PDFInfo
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- CN209974734U CN209974734U CN201920159192.9U CN201920159192U CN209974734U CN 209974734 U CN209974734 U CN 209974734U CN 201920159192 U CN201920159192 U CN 201920159192U CN 209974734 U CN209974734 U CN 209974734U
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Abstract
The utility model provides a micro-fluidic chip, including substrate and cover plate, substrate surface one end is equipped with sheath liquid entry and cell suspension entry in proper order, substrate surface middle part is equipped with two continuous phase oil entry relatively, the substrate surface other end is equipped with five liquid drop exports, all be connected with the microchannel on sheath liquid entry and the cell suspension entry, the microchannel that two continuous phase oil entry are connected forms the cross with parallel channel, the microchannel that connects out from the cross is the radiation and is divided into five separation channel and is connected with five liquid drop exports respectively, be equipped with liquid drop observation area on the microchannel that cross end and five separation channel's the end of assembling are connected between; two separation microelectrodes are oppositely arranged on two sides of the convergence end of the five separation channels. The utility model also provides a system for use the multiple cell of separation of micro-fluidic chip. The utility model discloses can improve cell sorting efficiency and reduce cost.
Description
Technical Field
The utility model relates to a system of micro-fluidic chip and multiple cell of separation.
Background
Flow cytometry plays an important role in cell sorting and analysis. It is the most widely used means for cell identification and separation at present, but it is expensive. The separation flux of the laser optical tweezers for sorting cells is low, and the system platform construction cost is very high. The research on the cell biological process from the single cell level has important significance, can essentially reveal the pathogenesis of the cancer, understand the principle of the small differentiation and the tissue development and identify the gene expression characteristic and the cell characteristic. Therefore, efficient and cost-effective differentiation of different cells from blood has become a popular direction of research.
SUMMERY OF THE UTILITY MODEL
In view of the above, the present invention provides a microfluidic chip to improve cell sorting efficiency and reduce cost.
The utility model relates to a technical solution:
a micro-fluidic chip comprises a substrate and a cover plate, wherein one end of the surface of the substrate is sequentially provided with a sheath fluid inlet and a cell suspension inlet, the middle part of the surface of the substrate is oppositely provided with two continuous phase oil inlets, the other end of the surface of the substrate is provided with five droplet outlets, the sheath fluid inlet and the cell suspension inlet are both connected with micro-channels, the micro-channel connected from the sheath fluid inlet is divided into two branches which are connected in parallel to two sides of the micro-channel of the cell suspension inlet, the micro-channel of the sheath fluid inlet and the micro-channel of the cell suspension inlet are combined to form a parallel channel, the micro-channel connected with the two continuous phase oil inlets and the parallel channel form a cross, the micro-channel connected from the cross is radially divided into five separation channels which are respectively connected with the five droplet outlets, and a droplet observation area is arranged on the micro-channel connected between the cross end, the micro-channels of the liquid drop observation area form a multi-channel structure; two sides of the convergence end of the five separation channels are oppositely provided with two separation microelectrodes, and the surface of the substrate provided with the microchannel is bonded with the cover plate to form the microfluidic chip.
Further, the width of the microchannel is 80 to 380 micrometers.
Further, the depth of the microchannel is 80 to 100 micrometers.
Further, the substrate is made of PDMA, PDMS, COC, acrylic plate or PMMA, and the cover plate is made of glass, PDMS, PMMA or PC.
Furthermore, the separation microelectrode is of a V-shaped structure, and two extending ends of the V-shaped structure are provided with electric connection points.
A system for separating a plurality of cells uses the microfluidic chip and comprises four injection pumps, a fluorescence detector, a computer and a programmable direct current power supply, wherein the four injection pumps are respectively connected with a sheath fluid inlet, a cell suspension inlet and two continuous phase oil inlets so as to inject corresponding liquid into the corresponding inlets; the programmable direct current power supply is electrically connected with the two separation microelectrodes, and the programmable direct current power supply has five different working modes aiming at five cells and outputs five pulse voltages so as to separate various cells; the computer is electrically connected with each injection pump, and the computer is provided with a preset program to control the injection speed of each injection pump.
Further, the fluorescence detector comprises a microscope and a CCD camera connected to the microscope.
Further, a cell collector is connected to each droplet outlet.
Further, the sheath fluid inlet, the cell suspension inlet, each continuous phase oil inlet and each liquid drop outlet are connected with a capillary glass tube and a micro-tube.
The micro-fluidic chip of the utility model adopts a cross-shaped liquid drop generating structure to realize the wrapping of cells and utilizes a method of sheath liquid focusing to arrange the cells orderly, thereby improving the success rate of single wrapping; in addition, the system for separating multiple cells effectively utilizes the phenomenon of liquid drop self-charging, realizes the separation of liquid drops wrapping the cells, and finally achieves the purpose of separating multiple cells; furthermore, the utility model provides a one set of simple feasible and be applicable to the sorting system of multiple cell, simultaneously, it has advantages such as efficient, weak point consuming time, low cost, pollution-free, application scope are wide.
Drawings
Fig. 1 is a schematic view of a flow channel structure of the microfluidic chip of the present invention;
FIG. 2 is a schematic diagram of a system for separating a plurality of cells.
Detailed Description
The technical solution in the embodiment of the present invention is clearly and completely described below with reference to the drawings in the embodiment of the present invention. It should be understood that the specific embodiments described herein are merely illustrative of the present invention and are not intended to limit the scope of the invention.
Referring to fig. 1, the present invention provides a microfluidic chip, which comprises a substrate and a cover plate, wherein one end of the surface of the substrate is sequentially provided with a sheath fluid inlet 11 and a cell suspension inlet 12, the middle part of the surface of the substrate is relatively provided with two continuous phase oil inlets 13, the other end of the surface of the substrate is provided with five droplet outlets 14, the sheath fluid inlet 11 and the cell suspension inlet 12 are both connected with microchannels 15, the microchannels 15 connected from the sheath fluid inlet 11 are divided into two branches which are connected in parallel to both sides of the microchannels 15 of the cell suspension inlet 12, the microchannels 15 of the sheath fluid inlet 11 and the microchannels 15 of the cell suspension inlet 12 are combined to form parallel channels 16, the microchannels 15 connected with the two continuous phase oil inlets 13 and the parallel channels 16 form a cross, the microchannels 15 connected from the cross are radially divided into five separation channels 17 which are respectively connected with the five droplet outlets 14, a liquid drop observation area 18 is arranged on the micro-channel 15 connected between the cross end and the convergence end of the five separation channels 17, and the micro-channel 15 of the liquid drop observation area 18 forms a multi-channel structure to reduce the speed of generating liquid drops, so that different cells form differential flow in the liquid drop observation area 18, and the cells with different sizes can be ensured to be preliminarily separated; two separation microelectrodes 19 are oppositely arranged on two sides of the convergence end of the five separation channels 17, so that five cells in the liquid drop can be deflected into the corresponding separation channels 17 under the action of voltage; the surface of the substrate provided with the micro-channel 15 is bonded with the cover plate to form the micro-fluidic chip.
The width of the microchannel 15 is 80 to 380 micrometers and the depth is 80 to 100 micrometers.
The substrate can be PDMA, PDMS, COC, acrylic plate or PMMA, and the cover plate can be glass, PDMS, PMMA or PC. In this example, the base sheet was made of PDMA and the cover sheet was made of glass.
The separation microelectrode 19 is in a V-shaped configuration with electrical connection points formed at the two protruding ends of the V-shaped configuration to provide a stable electrical connection.
Referring to fig. 2, the present invention further provides a system for separating multiple cells using the above microfluidic chip, the system includes four injection pumps 3, a fluorescence detector 4, a computer 5 and a programmable dc power supply 6, the four injection pumps 3 are respectively connected to a sheath fluid inlet 11, a cell suspension inlet 12 and two continuous phase oil inlets 13 to inject corresponding liquid into corresponding inlets, the fluorescence detector 4 and the programmable dc power supply 6 are respectively electrically connected to the computer 5, the fluorescence detector 4 is used for collecting fluorescence signals of droplets and transmitting the fluorescence signals to the computer 5, the computer 5 compares the received signals with the set signals to determine the type of cells wrapped by the droplets; the programmable direct current power supply 6 is electrically connected with the two separation microelectrodes 19, and the programmable direct current power supply 6 has five different working modes aiming at five cells and outputs five pulse voltages so as to separate various cells; the computer 5 is electrically connected with each injection pump 3, and the computer 5 is provided with a preset program to control the injection speed of each injection pump 3.
It will be understood by those skilled in the art that the fluorescence detector 4 comprises a microscope and a CCD camera attached to the microscope.
The system further comprises a cell collector 7 connected to each droplet outlet 14 for collecting the separated cells.
The sheath fluid inlet 11, the cell suspension inlet 12, each continuous phase oil inlet 13 and each droplet outlet 14 are connected with capillary glass tubes and micro-tubes and fixed by AB glue for connecting the injection pump 3 and the cell collector 7.
When in use, the handle is 2 multiplied by 106Cells were placed in 1ml pbs buffer and fluorescent stained, then, they were stained at 1: mixing 1 proportion and 2% sodium alginate solution, injecting sample from cell suspension inlet 12 at 3 μ l/h with syringe pump 3; at the same time, 1% sodium alginate solution8 mul/h sample is injected from a sheath fluid inlet 11; feeding the oil phase from two continuous phase oil inlets 13 at a rate of 190 μ l/h, and forming water-in-oil droplets of 100 μm by shearing the oil phase; in a droplet observation area 18 of the microfluidic chip, the droplet is decelerated and fluorescence is excited by a fluorescence detector 4, and a fluorescence signal is transmitted to a computer 5 for signal analysis; the computer 5 will send the processed signal to the programmable dc power supply 6; the programmable direct current power supply 6 outputs a pulse signal with a proper waveform according to the signal, so that the motion state of the liquid drop wrapping the cell is changed, and the sorted liquid drop is discharged from the liquid drop outlet 14 of the microfluidic chip and enters the cell collector 7.
The micro-fluidic chip of the utility model adopts a cross-shaped liquid drop generating structure to realize the wrapping of cells and utilizes a method of sheath liquid focusing to arrange the cells orderly, thereby improving the success rate of single wrapping; in addition, the system for separating multiple cells effectively utilizes the phenomenon of liquid drop self-charging, realizes the separation of liquid drops wrapping the cells, and finally achieves the purpose of separating multiple cells; furthermore, the utility model provides a one set of simple feasible and be applicable to the sorting system of multiple cell, simultaneously, it has advantages such as efficient, weak point consuming time, low cost, pollution-free, application scope are wide.
The above only is the embodiment of the present invention, not limiting the patent scope of the present invention, all utilize the equivalent structure or equivalent flow transformation that the content of the specification does, or directly or indirectly use in other related technical fields, all including in the same way the patent protection scope of the present invention.
Claims (9)
1. A micro-fluidic chip comprises a substrate and a cover plate, and is characterized in that one end of the surface of the substrate is sequentially provided with a sheath fluid inlet (11) and a cell suspension inlet (12), the middle part of the surface of the substrate is oppositely provided with two continuous phase oil inlets (13), the other end of the surface of the substrate is provided with five droplet outlets (14), the sheath fluid inlet (11) and the cell suspension inlet (12) are both connected with microchannels (15), the microchannels (15) connected from the sheath fluid inlet (11) are divided into two branches which are connected to two sides of the microchannels (15) of the cell suspension inlet (12) in parallel, the microchannels (15) of the sheath fluid inlet (11) and the microchannels (15) of the cell suspension inlet (12) are combined to form parallel channels (16), the microchannels (15) connected with the two continuous phase oil inlets (13) form a cross, and the microchannels (15) connected from the cross are radially divided into five separation channels (17) which are respectively connected with the five droplet inlets (12) The outlets (14) are connected, a liquid drop observation area (18) is arranged on the micro-channel (15) connected between the cross end and the convergence end of the five separation channels (17), and the micro-channel (15) of the liquid drop observation area (18) forms a multi-channel structure; two separation microelectrodes (19) are oppositely arranged on two sides of the convergence end of the five separation channels (17), and the surface of the substrate provided with the microchannel (15) is bonded with the cover plate to form the microfluidic chip.
2. The microfluidic chip according to claim 1, wherein the width of the microchannel (15) is 80 to 380 μm.
3. Microfluidic chip according to claim 1, characterized in that the depth of the microchannel (15) is 80 to 100 microns.
4. The microfluidic chip according to claim 1, wherein the substrate is PDMA, PDMS, COC, acrylic plate or PMMA, and the cover is glass, PDMS, PMMA or PC.
5. The microfluidic chip according to claim 1, wherein the separation microelectrode (19) is in a V-shaped configuration, and electrical connection points are formed at two extending ends of the V-shaped configuration.
6. A system for separating a plurality of cells, which is characterized in that the microfluidic chip of any one of claims 1 to 5 is used, the system comprises four injection pumps (3), a fluorescence detector (4), a computer (5) and a programmable direct current power supply (6), the four injection pumps (3) are respectively connected with a sheath fluid inlet (11), a cell suspension inlet (12) and two continuous phase oil inlets (13) so as to inject corresponding liquid into the corresponding inlets, the fluorescence detector (4) and the programmable direct current power supply (6) are respectively electrically connected with the computer (5), the fluorescence detector (4) is used for collecting fluorescence signals of liquid drops and transmitting the fluorescence signals to the computer (5), and the computer (5) judges the type of the cells wrapped by the liquid drops by comparing the received signals with set signals; the programmable direct current power supply (6) is electrically connected with the two separation microelectrodes (19), and the programmable direct current power supply (6) outputs five pulse voltages aiming at five different working modes of five cells so as to separate various cells; the computer (5) is electrically connected with each injection pump (3), and the computer (5) is provided with a preset program to control the injection speed of each injection pump (3).
7. The system for separating a plurality of cells according to claim 6, wherein the fluorescence detector (4) comprises a microscope and a CCD camera connected to the microscope.
8. The system for separating a plurality of cells according to claim 6, wherein a cell collector (7) is connected to each droplet outlet (14).
9. The system for separating a plurality of cells according to claim 6, wherein a capillary glass tube and a micro tube are connected to the sheath fluid inlet (11), the cell suspension inlet (12), each continuous phase oil inlet (13) and each droplet outlet (14).
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| Application Number | Priority Date | Filing Date | Title |
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| CN201920159192.9U CN209974734U (en) | 2019-01-28 | 2019-01-28 | Micro-fluidic chip and system for separating various cells |
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| CN201920159192.9U CN209974734U (en) | 2019-01-28 | 2019-01-28 | Micro-fluidic chip and system for separating various cells |
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Effective date of registration: 20210723 Address after: 519000 room 202-4, floor 2, building 20, science and Innovation Park, Gangwan 1, Jintang Road, Tangjiawan Town, high tech Zone, Zhuhai, Guangdong Patentee after: Zhuhai yinfangzhou biochip Co.,Ltd. Address before: 528225 room a401-11, Nanhai Software Technology Park (building a, R & D building), Shishan town, Nanhai District, Foshan City, Guangdong Province Patentee before: FOSHAN GEWEI TECHNOLOGY Co.,Ltd. |
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