CN107699485A - Microelectrode fluidic chip and adjustable parameter single cell electroporation device - Google Patents

Microelectrode fluidic chip and adjustable parameter single cell electroporation device Download PDF

Info

Publication number
CN107699485A
CN107699485A CN201711082795.5A CN201711082795A CN107699485A CN 107699485 A CN107699485 A CN 107699485A CN 201711082795 A CN201711082795 A CN 201711082795A CN 107699485 A CN107699485 A CN 107699485A
Authority
CN
China
Prior art keywords
microelectrode
cell
electroporation
fluidic chip
fluidic
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CN201711082795.5A
Other languages
Chinese (zh)
Other versions
CN107699485B (en
Inventor
朱真
耿杨烨
潘任豪
王颖瀛
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Southeast University
Original Assignee
Southeast University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Southeast University filed Critical Southeast University
Priority to CN201711082795.5A priority Critical patent/CN107699485B/en
Publication of CN107699485A publication Critical patent/CN107699485A/en
Application granted granted Critical
Publication of CN107699485B publication Critical patent/CN107699485B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12MAPPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00Constructional details, e.g. recesses, hinges
    • C12M23/02Form or structure of the vessel
    • C12M23/16Microfluidic devices; Capillary tubes
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12MAPPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M35/00Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
    • C12M35/02Electrical or electromagnetic means, e.g. for electroporation or for cell fusion

Abstract

The invention discloses a kind of microelectrode fluidic chip and adjustable parameter single cell electroporation device, microelectrode fluidic chip includes transparency carrier and microfluidic channel layer, microfluidic channel layer is located above transparency carrier, microelectrode is integrated with transparency carrier, microelectrode includes interdigital electrode and parallel pole, interdigital electrode is with parallel distribution of electrodes in the lower section of micro-fluidic electroporation passage, and interdigital electrode and parallel pole are sequentially distributed from cell suspending liquid entrance to cell outlet.Adjustable parameter single cell electroporation device includes power module of voltage regulation, module, PCB control circuits and digital display module, chip carrier and microelectrode fluidic chip occur for function signal.The present invention can realize carries out electroporation processing successively to individual cells;Reduce the cellular damage even death rate;Improve cell electrotransfection efficiency;The species and quantity of access electrode are selected by control circuit, changes the frequency, amplitude, dutycycle of electric signal, the optimization of Electroporation parameters is realized for different cell lines.

Description

Microelectrode fluidic chip and adjustable parameter single cell electroporation device
Technical field
The invention belongs to microflow control technique and single cell electroporation, electrotransfection field, more particularly to one kind to be based on microelectrode The adjustable parameter single cell electroporation device of fluidic chip.
Background technology
Microflow control technique (Microfluidics) is in a few micrometers to hundreds of microns of fluid channel processing or behaviour using size Vertical volume is the Science and Technology involved by nanoliter system for arriving microlitre fluid, is one and is related to micro-nano technology, physics, micro- electricity The emerging cross discipline in the fields such as sub-, biology, chemistry, new material.Based on the chip of microflow control technique to be miniaturized, integrate Feature is turned to, generally again by title chip lab (Lab on a Chip) and micro-total analysis system (microTAS).Micro-fluidic skill Art is considered as having huge development potentiality in biomedical research and is widely applied prospect.
, it is necessary to apply the electricity in the external world, optical signalling when carrying out biological, chemical sample detection and analysis with micro-fluidic chip Excitation reflects the parameter of sample to be tested by output electricity, optical signalling.Especially, for electrical detection analytical technology For, it need to utilize micro-processing technology Integrated electrode on micro-fluidic chip, be interacted with realizing with outer signals, so that in miniflow Control and electrical detection method is integrated on chip.The micro-fluidic chip of integrated microelectrode possesses many superior performances:Detected in manipulation While biological specimen, electric field caused by microelectrode can promote cell to produce some physiological reactions, such as perforate, crack, or will Associated electrical information (such as resistance antinoise signal) feeds back to experimental system to realize cell detection, and possesses high sensitivity, response The advantages that fast and the potentiality of device miniaturization.
Cell electroporation technology (Electroporation) is also known as electrotransfection technology, is commonly used in cell transfection technique A kind of approach.Because cell membrane has selective penetrated property to external substance, control gene of eucaryote cell experiment needs thin to eucaryon Specific biological DNA, RNA fragment of born of the same parents' input.Apply the electrical potential difference of some strength in cell membrane both sides and continue for some time, carefully Micropore just can be produced on after birth, strengthens membrane passage.When electroporation occurs for cell membrane, its permeability and membrane conductance meeting Instantaneous increase, so that hydrophilic molecules, DNA, protein, virion, drug particles etc. can not pass through cell under normal circumstances The molecule of film gets enter into cell.After removing electrical potential difference in a short time, cell membrane can self-recovery, again turn into selectivity Permeability barrier.Compared with traditional chemical transfection and virus transfection, electroporation has more wide applicability and superiority:It is applicable In plasmid and tens KB genomic fragment, without chemical viral pollution, acellular permanent damage, transient transfection the advantages that.Cause This, electroporation technology has broad application prospects in fields such as biophysics, molecular biology, clinical medicine.
Electrode size is big in traditional cell electroporation instrument, electrode spacing still in macro-size, in ten mm-scales, And cell size is in micro-meter scale, therefore the voltage that need to apply is big, about hundreds of volts, and electric field is uneven, each cell Residing electric field environment is different, causes that the cell close to electrode is easily dead, and the cell at compared with weak electric field can not be worn again Hole transfects, and survival rate and transfection efficiency are all than relatively low.The parameters such as existing micro-fluidic electroporation device electrode spacing are fixed, experiment Operability and Electroporation parameters controllability by chip design limited, lack versatility.In addition, commercialized electricity The key parameters such as signal frequency, the amplitude of instrument are transfected all by factory settings, client can not optimize regulation, for special Cell, such as primary cell, immunocyte transfection efficiency be not high.
The content of the invention
Goal of the invention:In order to solve the problems, such as that prior art is present, electroporation device is set to be common to various cells, the present invention A kind of microelectrode fluidic chip and adjustable parameter single cell electroporation device is provided.
Technical scheme:A kind of microelectrode fluidic chip, including transparency carrier and microfluidic channel layer, microfluidic channel layer position Above transparency carrier, the microfluidic channel layer includes sheath inflow entrance, cell suspending liquid entrance, sheath circulation road, cell and suspended Liquid passage, micro-fluidic electroporation passage and cell outlet;Sheath inflow entrance connects with sheath circulation road, cell suspending liquid entrance and cell Suspension passage is connected, and the end of sheath circulation road is converged with cell suspending liquid channel end and with the one of micro-fluidic electroporation passage End connection, the other end of micro-fluidic electroporation passage connect with cell outlet;Microelectrode, micro- electricity are integrated with the transparency carrier Pole includes interdigital electrode and parallel pole, and interdigital electrode, in the lower section of micro-fluidic electroporation passage, and is pitched with parallel distribution of electrodes Refer to electrode and parallel pole is sequentially distributed from cell suspending liquid entrance to cell outlet;Microelectrode includes microelectrode exit, micro- Electrode leads to client is distributed in transparency carrier both sides.
Preferably, the interdigital electrode and/or parallel pole have multiple and uniform intervals to be distributed.Electric Field Distribution can be made more Add uniformly, make the electric field environment residing for each cell close, improve survival rate and transfection efficiency.
Preferably, the sheath circulation road includes two passage tributaries, and two passage tributaries are respectively from cell suspending liquid entrance Both sides connect around cell suspending liquid entrance with cell suspending liquid passage, and the sheath stream in two passage tributaries can be effectively by cell The flowing of microchannel center is converged in, realizes and electroporation processing is carried out successively to individual cells.
Preferably, in addition to cell suspending liquid input pipe, sheath stream input pipe, cell efferent duct, cell suspending liquid input pipe Connected with cell suspending liquid entrance, sheath stream input pipe connects with sheath inflow entrance, and cell efferent duct connects with cell delivery outlet.
Preferably, transparency carrier is provided with bonding cross alignment mark, and microfluidic channel layer is provided with being bonded with ten Four square alignment marks of the bonding that word alignment mark is engaged, bonding make transparency carrier be combined more with microfluidic channel layer Accurately, with ensure electrode be able to be located at micro-fluidic electroporation passage in.
Preferably, the finger spacing of interdigital electrode is 50~80 μm;The electrode spacing of parallel pole is 150~200 μm.
A kind of adjustable parameter single cell electroporation device using microelectrode fluidic chip, including power module of voltage regulation, letter Number signal generating module, PCB control circuits and digital display module, chip carrier and microelectrode fluidic chip, chip carrier are provided with Chip slot, the microelectrode fluidic chip are fixed in chip slot, and PCB control circuits and digital display module output end are visited for spring Pin, the spring probe make electrical contact with microelectrode exit;Power module of voltage regulation is used to module, PCB controls occur for function signal Circuit processed and digital display module and the power supply of microelectrode fluidic chip;Function signal occurs module and is used to believe needed for output power supply perforation Number.
Preferably, the PCB control circuits and digital display module include on-off circuit and amplifier, and on-off circuit is defeated by single channel Enter and be divided into multiple-channel output, amplifier is used for the amplitude for adjusting output signal;The input contiguous function signal of on-off circuit occurs The output end of module, the input of the output end connection amplifier of on-off circuit;The output end connection microelectrode of amplifier is drawn End.
Preferably, amplifier is gain adjustable amplifier.
Preferably, interdigital electric field of the interdigital electrode is more than the electric field between electrodes of parallel pole.
Beneficial effect:A kind of microelectrode fluidic chip provided by the invention, compare prior art, microelectrode fluidic chip In microelectrode include interdigital electrode and parallel pole, interdigital electrode is located at micro-fluidic electroporation passage upstream, produces highfield, Cell suspending liquid by when by electric field action, electroporation occurs for cell membrane;Parallel pole is located under micro-fluidic electroporation passage Trip, applies and keeps existing fringing field, can maintain cell electroporation state, strengthen and improve the transfection efficiency of cell.Each cell Residing electric field environment is close, and cellular damage caused by reducing conventional electroporation instrument highfield and electric-field intensity difference is even dead Die.
Adjustable parameter single cell electroporation device based on microelectrode fluidic chip, compare prior art, and 1) device For complete single cell electroporation experimental system, it is convenient to carry out single cell electroporation experiment, be placed on inverted microscope Observe the dynamic process of cell electroporation;2) device uses microelectrode fluidic chip, and spacing is short between electrode, to reach cell electricity The effect of perforation and the voltage that applies is low in 10V magnitudes, cell mortality, safe operation;3) device to individual cells successively Electroporation is carried out, and the electric field environment residing for each cell is close, reduces cell caused by uncontrollable factor in experimentation and damages Wound;4) electrode geometrical parameter of the core microelectrode fluidic chip of the device can be designed according to requirement of experiment, and electricity is controlled by PCB Road selects the species of electrode used in electroporation and number and selects waveform, frequency, dutycycle and the amplitude of electric signal, Realize the optimization regulation of electroporation experiment parameter;5) the microelectrode fluidic chip of the device designs simultaneously in microfluidic channel downstream Parallel pole is machined with, existing fringing field is produced when cell flows through the region, effectively enhances cell electrotransfection efficiency.
Brief description of the drawings
Fig. 1 is the three dimensional structure diagram of microelectrode fluidic chip in the present invention;
Fig. 2 is the three dimensional structure diagram of microfluidic channel layer in microelectrode fluidic chip in the present invention;
Fig. 3 is the transparency carrier three dimensional structure diagram for integrating microelectrode in the present invention in microelectrode fluidic chip;
Fig. 4 is microelectrode fluidic chip A-A profile structure diagram;
Fig. 5 is the structured flowchart of the adjustable parameter single cell electroporation device of the present invention.
Embodiment
The invention will be further described with specific embodiment below in conjunction with the accompanying drawings.
As shown in figure 1, microelectrode fluidic chip includes transparency carrier 10 and microfluidic channel layer 20, microfluidic channel layer 20 Above transparency carrier 10.As shown in Fig. 2 the microfluidic channel layer 20 includes sheath inflow entrance 201, cell suspending liquid entrance 202nd, sheath circulation road 203, cell suspending liquid passage 204, micro-fluidic electroporation passage 205 and cell outlet 206;Sheath inflow entrance 201 connect with sheath circulation road 203, and cell suspending liquid entrance 202 connects with cell suspending liquid passage 204, the end of sheath circulation road 203 End is converged with the end of cell suspending liquid passage 204 and connected with one end of micro-fluidic electroporation passage 205, and micro-fluidic electroporation leads to The other end in road 205 connects with cell outlet 206.As shown in figure 3, it is integrated with microelectrode, microelectrode on the transparency carrier 10 Including interdigital electrode 101 and parallel pole 102, interdigital electrode 101 and parallel pole 102 are distributed in micro-fluidic electroporation passage 205 lower section, micro-fluidic electroporation passage 205 is just from interdigital electrode 101 and parallel pole 102 from the point of view of vertical view Between pass through, the two-stage of electrode is located at the micro-fluidic both ends of electroporation passage 205 respectively, and interdigital electrode 101 and parallel pole 102 from Cell suspending liquid entrance 202 is sequentially distributed to cell outlet 206, i.e. interdigital electrode 101 close to cell suspending liquid entrance 202, Upstream;Parallel pole 102 is close to cell outlet 206, in downstream;Microelectrode includes microelectrode exit 103, microelectrode exit 103 are distributed in the both sides of transparency carrier 10.
Interdigital electrode 101 in the present embodiment has three, and parallel pole 102 has one, can actually be arranged as required to Corresponding number, parallel pole 102 can also have multiple.Interval is uniformly distributed between multiple microelectrodes, designs phase as needed The spacing of adjacent microelectrode, Electric Field Distribution can be made more uniform, make the electric field environment residing for each cell close, improve survival rate And transfection efficiency.Microelectrode spacing is embodied in geometric electrode size and arrangement when making transparency carrier 10 and designed.
The sheath circulation road 203 includes two passage tributaries, and two passage tributaries are respectively from 202 liang of cell suspending liquid entrance Side connects around cell suspending liquid entrance 202 with cell suspending liquid passage 204, the sheath stream in two passage tributaries can effectively by Cell converges in the flowing of microchannel center, realizes and carries out electroporation processing successively to individual cells.
As shown in figure 4, also include sheath stream input pipe 301, cell suspending liquid input pipe 302, cell efferent duct 303, cell Suspension input pipe 302 connects with cell suspending liquid entrance 202, and sheath stream input pipe 301 connects with sheath inflow entrance 201, and cell is defeated Outlet pipe 303 connects with cell delivery outlet 206, to facilitate cell suspending liquid, the injection of sheath stream and the outflow of mixing liquid.Sheath stream Input pipe 301, cell suspending liquid input pipe 302, cell efferent duct 303 typically use teflon pipe, it is possible to use stainless steel tube, Specific diameter can determine according to the cell suspending liquid entrance 202, sheath inflow entrance 201, the aperture of cell outlet 206.
Transparency carrier 10 is provided with bonding cross alignment mark 104, and microfluidic channel layer 20 is provided with being bonded with ten Four square alignment marks 207 of the bonding that word alignment mark 104 is engaged, i.e., can also using sphere of movements for the elephants shape alignment mark It is the same using the alignment mark of other shapes, effect.Sheath inflow entrance 201, cell suspending liquid entrance are additionally provided with transparency carrier 10 202nd, the processing alignment mark 105 of cell outlet 206.Bonding shares four directions with lucky sunk key at cross-shaped alignment marks 104 In shape alignment mark 207, transparency carrier 10 is set to be combined with microfluidic channel layer 20 more accurate, to ensure that micro-fluidic electroporation leads to Road 205 be able to be located in microelectrode.In the occasion without fine registration, bonding alignment mark can not be also designed.
Spacing between the two end electrodes of microelectrode can be designed as needed, the interdigital electrode 101 of the present embodiment Finger spacing be 50~80 μm;The electrode spacing of parallel pole 102 is 150~200 μm.Both changeabilities are embodied in transparent base Electrode parameter design when plate 10 designs.
As shown in figure 5, use the adjustable parameter single cell electroporation device of the microelectrode fluidic chip, including voltage-stabilized power supply Module, PCB control circuits and digital display module, chip carrier and microelectrode fluidic chip, chip carrier occur for module, function signal Chip slot is provided with, the microelectrode fluidic chip is fixed in chip slot, and PCB control circuits and digital display module output end are bullet Spring probe, the spring probe make electrical contact with microelectrode exit.
Power module of voltage regulation is used to module, PCB control circuits and digital display module and microelectrode stream control occur for function signal Chip power supply.
Module, which occurs, for function signal can control the parameters such as the waveform for being applied to microelectrode power on signal, frequency, dutycycle, For exporting perforation desired signal of powering.
PCB control circuits and digital display module can be applied to microelectrode power on signal by gain adjustable amplifier to adjust Amplitude, realize for different cell lines Electroporation parameters optimization regulation.The PCB control circuits and digital display module bag On-off circuit and amplifier are included, single channel input is divided into multiple-channel output by on-off circuit;Amplifier is used for the width for adjusting output signal Value;The output end of the input contiguous function signal generating module of on-off circuit, the output end connection amplifier of on-off circuit Input;The output end connection microelectrode exit of amplifier.Because microelectrode have it is multiple, amplifier also have it is multiple, often The individual signal from on-off circuit output is accessed corresponding microelectrode after amplifier amplifies.Amplifier amplifies for adjustable gain Device, gain that can be as needed to each amplifier are adjusted, so as to realize the signal width being applied on each microelectrode Value is different.
Voltage stabilizing circuit module can be used the Switching Power Supply voltage stabilizing chip such as LM2575, MAX1715, efficiency high, small power consumption, specifically Chip can be chip used according to PCB control circuits and digital display module supply voltage determine;Module, which occurs, for function signal to make Chip MAX038 occurs with single chip integrated function to complete, high-frequency high-precision output waveform can be produced, output waveform distortion is small, drift Small, wide frequency range is moved, the waveforms such as sine wave, square wave can be produced, and frequency, dutycycle are adjustable;Make in PCB control circuits Realize that electric signal single channel inputs multiple-channel output with on-off circuit, adjustable gain function is realized using gain adjustable amplifier. In the present embodiment, there are three groups of interdigital electrodes and one group of parallel pole, therefore PCB control circuits are that single channel inputs the output of four tunnels, are increased Benefit is respectively set to 0 times, 1 times, 2 times and 3 times.
The break-make of microelectrode power on signal is by the chip in PCB control circuit modules and switch co- controlling, chip The input of single channel electric signal, the output of multi-channel electric signal are realized, the species sum of electrode is accessed by PCB control circuits switch-mode regulation Amount, the gating of multi-electrode is realized, change the time that cell is exposed in electric field, so as to control the work bar of cell electroporation Part.
Interdigital electric field of the interdigital electrode 101 is more than the electric field between electrodes of parallel pole 102, when cell passes through upstream When, electroporation can be implemented to cell;Cell after electroporation continues downward downstream, the electric field between electrodes of parallel pole 102 It is relatively low, cell electroporation state can be maintained, strengthens and improves the transfection efficiency of cell.
The making of microelectrode fluidic chip and be mounted to adjustable parameter single cell electroporation device process it is as follows:
(1) gold microelectrode is made using lift-off techniques on 4 cun of transparency carriers;Utilize and be based in 4 cun of silicon wafers The soft light carving technology of SU-8 photoresists makes PDMS microfluidic channel layers;Glass, (poly-methyl methacrylate can be used in transparency carrier Fat) transparent insulation material such as PMMA makes;The precious metal materials such as gold, platinum can be used by the technique such as electroplating or depositing in microelectrode Make.Transparency carrier material ensure that can carry out optical microphotograph observation in experimentation;The materials chemistry such as gold, platinum strong inert, Good conductivity, no biotoxicity.
(2) as shown in Figure 1 to 4, under the observation of stereomicroscope, punched on PDMS microfluidic channel layers, To process cell suspending liquid entrance, sheath inflow entrance, cell outlet.
(3) transparency carrier is cleaned, dried up respectively with PDMS microfluidic channels layer, is subsequently placed in oxygen plasma cleaning Permanent bonding is realized after carrying out surface modification treatment in machine.By on PDMS microfluidic channel layers under stereoscopic sem observation during bonding Bonding four square alignment marks and being bonded with cross alignment mark center superposition on transparency carrier, to ensure that electrode is proper It can be located in micro-fluidic electroporation passage.Then as shown in figure 4, inserting cell suspending liquid input on microelectrode fluidic chip Pipe, sheath stream input pipe, cell efferent duct.
(4) as shown in figure 5, microelectrode fluidic chip is fixed in the chip slot in chip carrier, by PCB control circuits Module and chip carrier fastened by screw, the spring probe and microelectrode for making control circuit realize electrical contact under stress, then Connect power module of voltage regulation and module occurs for function signal, form the unicellular electricity of the adjustable parameter based on microelectrode fluidic chip and wear Aperture apparatus.

Claims (10)

1. a kind of microelectrode fluidic chip, it is characterised in that including transparency carrier (10) and microfluidic channel layer (20), microfluid Channel layer (20) is located above transparency carrier (10), and the microfluidic channel layer (20) includes sheath inflow entrance (201), cell suspends Liquid entrance (202), sheath circulation road (203), cell suspending liquid passage (204), micro-fluidic electroporation passage (205) and cell outlet (206);Sheath inflow entrance (201) connects with sheath circulation road (203), cell suspending liquid entrance (202) and cell suspending liquid passage (204) connect, the end of sheath circulation road (203) is converged with cell suspending liquid passage (204) end and led to micro-fluidic electroporation One end connection in road (205), the other end of micro-fluidic electroporation passage (205) connect with cell outlet (206);The transparent base Microelectrode is integrated with plate (10), microelectrode includes interdigital electrode (101) and parallel pole (102), interdigital electrode (101) peace Row electrode (102) is distributed in the lower section of micro-fluidic electroporation passage (205), and interdigital electrode (101) and parallel pole (102) from Cell suspending liquid entrance (202) is sequentially distributed to cell outlet (206);Microelectrode includes microelectrode exit (103), microelectrode Exit (103) is distributed in transparency carrier (10) both sides.
2. microelectrode fluidic chip according to claim 1, it is characterised in that the interdigital electrode (101) and/or parallel Electrode (102) has multiple and uniform intervals to be distributed.
3. microelectrode fluidic chip according to claim 1, it is characterised in that the sheath circulation road (203) includes two Passage tributary, two passage tributaries respectively from cell suspending liquid entrance (202) both sides around cell suspending liquid entrance (202) with it is thin Born of the same parents' suspension passage (204) connects.
4. microelectrode fluidic chip according to claim 1, it is characterised in that also including sheath stream input pipe (301), cell Suspension input pipe (302), cell efferent duct (303), cell suspending liquid input pipe (302) and cell suspending liquid entrance (202) Connection, sheath stream input pipe (301) connect with sheath inflow entrance (201), and cell efferent duct (303) connects with cell delivery outlet (206).
5. microelectrode fluidic chip according to claim 1, it is characterised in that transparency carrier (10) is provided with bonding with ten Word alignment mark (104), microfluidic channel layer (20) are provided with the bonding being engaged with bonding with cross alignment mark (104) and used Four square alignment marks (207).
6. microelectrode fluidic chip according to claim 1, it is characterised in that the finger spacing of interdigital electrode (101) is 50 ~80 μm;The electrode spacing of parallel pole (102) is 150~200 μm.
7. a kind of adjustable parameter single cell electroporation device using the microelectrode fluidic chip as described in claim 1-5, it is special Sign is, including module, PCB control circuits and digital display module, chip carrier and micro- electricity occur for power module of voltage regulation, function signal Pole fluidic chip, chip carrier are provided with chip slot, and the microelectrode fluidic chip is fixed in chip slot, PCB control circuits And digital display module output end is spring probe, the spring probe makes electrical contact with microelectrode exit (103);Power module of voltage regulation For module, PCB control circuits and digital display module and the power supply of microelectrode fluidic chip to occur for function signal;Function signal occurs Module is used to export power supply perforation desired signal.
8. adjustable parameter single cell electroporation device according to claim 6, it is characterised in that the PCB control circuits And digital display module includes on-off circuit and amplifier, single channel input is divided into multiple-channel output by on-off circuit, and amplifier is used to adjust The amplitude of output signal;The output end of the input contiguous function signal generating module of on-off circuit, the output end of on-off circuit Connect the input of amplifier;The output end connection microelectrode exit of amplifier.
9. adjustable parameter single cell electroporation device according to claim 7, it is characterised in that amplifier is adjustable gain Amplifier.
10. adjustable parameter single cell electroporation device according to claim 7, it is characterised in that the interdigital electrode (101) interdigital electric field is more than the electric field between electrodes of parallel pole (102).
CN201711082795.5A 2017-11-06 2017-11-06 Microelectrode flow control chip and parameter-adjustable single-cell electroporation device Active CN107699485B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201711082795.5A CN107699485B (en) 2017-11-06 2017-11-06 Microelectrode flow control chip and parameter-adjustable single-cell electroporation device

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201711082795.5A CN107699485B (en) 2017-11-06 2017-11-06 Microelectrode flow control chip and parameter-adjustable single-cell electroporation device

Publications (2)

Publication Number Publication Date
CN107699485A true CN107699485A (en) 2018-02-16
CN107699485B CN107699485B (en) 2020-08-11

Family

ID=61178226

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201711082795.5A Active CN107699485B (en) 2017-11-06 2017-11-06 Microelectrode flow control chip and parameter-adjustable single-cell electroporation device

Country Status (1)

Country Link
CN (1) CN107699485B (en)

Cited By (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108982335A (en) * 2018-06-29 2018-12-11 浙江大学 A kind of suspension cell counting chip based on electrical impedance and preparation method thereof and method of counting
CN109239381A (en) * 2018-08-02 2019-01-18 深圳大学 A kind of micro-fluidic chip and micro fluidic device for biomolecule detection
WO2019183998A1 (en) * 2018-03-28 2019-10-03 东南大学 Micro-flow control chip for cell tissue culture and real-time monitoring, and use method therefor
CN110632138A (en) * 2019-11-01 2019-12-31 江南大学 Interdigital electrode chip
CN111257378A (en) * 2020-02-24 2020-06-09 东南大学 Passive wireless sensing detection device
CN111774115A (en) * 2020-05-29 2020-10-16 东南大学 Clamp device for fixing micro-fluidic chip with electrodes
CN113000078A (en) * 2019-12-19 2021-06-22 深圳华大生命科学研究院 Chip and preparation method thereof
CN113166706A (en) * 2018-08-31 2021-07-23 查尔斯·斯塔克·德雷珀实验室有限公司 High-throughput efficient cell transfection method and device
CN113528332A (en) * 2021-07-19 2021-10-22 中山大学 Intracellular and extracellular electrophysiological recording system and method for automatic electroporation regulation and screening
CN114870917A (en) * 2022-05-09 2022-08-09 南京大学 Microfluidic chip for identifying different cells and preparation method and detection platform thereof
CN114940973A (en) * 2021-08-27 2022-08-26 南京微欣利康科技有限公司 Microfluidic cell sorting device, sorting method of tumor stem cells and screening method of anti-cancer drugs
EP4100506A4 (en) * 2020-04-07 2023-05-03 Hewlett-Packard Development Company, L.P. Microfluidic chip cell sorting and transfection
WO2024015072A1 (en) * 2022-07-15 2024-01-18 Hewlett-Packard Development Company, L.P. Cell porating and optically detecting microfluidic devices

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101923053A (en) * 2010-07-19 2010-12-22 杭州师范大学 Device and method for continuously analyzing single-cell contents by miniflow control chip at high speed
CN106085845A (en) * 2016-07-12 2016-11-09 重庆大学 Cell electroporation chip apparatus based on U-shaped groove microelectrode array and processing method thereof
CN106148187A (en) * 2016-07-20 2016-11-23 国家纳米科学中心 For expressing unicellular sorting and the micro-fluidic chip of polygenic locus detection of EGFR
CN106591118A (en) * 2017-01-05 2017-04-26 博奥生物集团有限公司 Cell in-situ electroporation device and use method
CN106676001A (en) * 2016-12-29 2017-05-17 中国科学院微电子研究所 Selectively persistent-flow cell electroporation system and method based on cross narrow channel

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101923053A (en) * 2010-07-19 2010-12-22 杭州师范大学 Device and method for continuously analyzing single-cell contents by miniflow control chip at high speed
CN106085845A (en) * 2016-07-12 2016-11-09 重庆大学 Cell electroporation chip apparatus based on U-shaped groove microelectrode array and processing method thereof
CN106148187A (en) * 2016-07-20 2016-11-23 国家纳米科学中心 For expressing unicellular sorting and the micro-fluidic chip of polygenic locus detection of EGFR
CN106676001A (en) * 2016-12-29 2017-05-17 中国科学院微电子研究所 Selectively persistent-flow cell electroporation system and method based on cross narrow channel
CN106591118A (en) * 2017-01-05 2017-04-26 博奥生物集团有限公司 Cell in-situ electroporation device and use method

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
KANG ET AL: "Microfluidic device for stem cell differentiation and localized electroporation of postmitotic neurons", 《LAB ON A CHIP》 *

Cited By (17)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2019183998A1 (en) * 2018-03-28 2019-10-03 东南大学 Micro-flow control chip for cell tissue culture and real-time monitoring, and use method therefor
CN108982335A (en) * 2018-06-29 2018-12-11 浙江大学 A kind of suspension cell counting chip based on electrical impedance and preparation method thereof and method of counting
CN109239381A (en) * 2018-08-02 2019-01-18 深圳大学 A kind of micro-fluidic chip and micro fluidic device for biomolecule detection
CN113166706A (en) * 2018-08-31 2021-07-23 查尔斯·斯塔克·德雷珀实验室有限公司 High-throughput efficient cell transfection method and device
CN110632138A (en) * 2019-11-01 2019-12-31 江南大学 Interdigital electrode chip
CN113000078A (en) * 2019-12-19 2021-06-22 深圳华大生命科学研究院 Chip and preparation method thereof
WO2021169019A1 (en) * 2020-02-24 2021-09-02 东南大学 Passive wireless sensor detecting discrete droplets and bubbles
CN111257378A (en) * 2020-02-24 2020-06-09 东南大学 Passive wireless sensing detection device
US11408845B2 (en) 2020-02-24 2022-08-09 Southeast University Passive wireless sensor for detecting discrete droplets and bubbles
EP4100506A4 (en) * 2020-04-07 2023-05-03 Hewlett-Packard Development Company, L.P. Microfluidic chip cell sorting and transfection
CN111774115A (en) * 2020-05-29 2020-10-16 东南大学 Clamp device for fixing micro-fluidic chip with electrodes
CN113528332A (en) * 2021-07-19 2021-10-22 中山大学 Intracellular and extracellular electrophysiological recording system and method for automatic electroporation regulation and screening
CN113528332B (en) * 2021-07-19 2023-10-24 中山大学 Intracellular and extracellular electrophysiological recording system and method for automatic electroporation regulation screening
CN114940973A (en) * 2021-08-27 2022-08-26 南京微欣利康科技有限公司 Microfluidic cell sorting device, sorting method of tumor stem cells and screening method of anti-cancer drugs
CN114870917A (en) * 2022-05-09 2022-08-09 南京大学 Microfluidic chip for identifying different cells and preparation method and detection platform thereof
CN114870917B (en) * 2022-05-09 2023-06-06 南京大学 Microfluidic chip for identifying different cells, preparation method thereof and detection platform
WO2024015072A1 (en) * 2022-07-15 2024-01-18 Hewlett-Packard Development Company, L.P. Cell porating and optically detecting microfluidic devices

Also Published As

Publication number Publication date
CN107699485B (en) 2020-08-11

Similar Documents

Publication Publication Date Title
CN107699485A (en) Microelectrode fluidic chip and adjustable parameter single cell electroporation device
Kar et al. Single-cell electroporation: current trends, applications and future prospects
Wang et al. Single-cell electroporation
Chen et al. A simplified microfluidic device for particle separation with two consecutive steps: Induced charge electro-osmotic prefocusing and dielectrophoretic separation
Gomez Biological applications of microfluidics
Andersson et al. Microfluidic devices for cellomics: a review
US9995668B2 (en) Apparatus for manipulating, modifying and characterizing particles in a micro channel
WO2019183998A1 (en) Micro-flow control chip for cell tissue culture and real-time monitoring, and use method therefor
Guo et al. Controllable in-situ cell electroporation with cell positioning and impedance monitoring using micro electrode array
Tajik et al. Simple, cost-effective, and continuous 3D dielectrophoretic microchip for concentration and separation of bioparticles
BRPI0910898A2 (en) ex-vivo multi-dimensional system for the separation and isolation of cells, vesicles, nanoparticles and biomarkers
JP2002533081A (en) Microsystems for cell penetration and cell fusion
CN101928666A (en) Flow type electroporation device and system
CN101857836B (en) Flow electroporation device and system
KR20160133812A (en) Apparatus comprising nanoporous membrane for separating organic molecule
CN101870949B (en) Electroporated chip and porous plate device base on electroporated chip
CN110918139A (en) Microfluidic chip, device containing same and sample concentration method
CN109289951A (en) Drop breakup micro-fluidic chip and application
CN102174387A (en) Low-voltage direct-current controlled continuous flow cell electrofusion chip
CN110923111A (en) Microfluidic chip, device containing the same, and method for detecting or sorting sample
Lee et al. Continuous medium exchange and optically induced electroporation of cells in an integrated microfluidic system
Chen et al. Rapid concentration of nanoparticles with DC dielectrophoresis in focused electric fields
Wang et al. A two-stage separation of circulating tumor cells based on deterministic lateral displacement and dielectrophoresis techniques
He et al. Micro pulsed radio-frequency electroporation chips
CN108949497B (en) Specific single cell fixed-point capturing chip for trace circulating tumor cells

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant