CN102174387A - Low-voltage direct-current controlled continuous flow cell electrofusion chip - Google Patents
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Abstract
The invention provides a microelectrode array chip which can be used for continuous flow cell electrofusion. The chip consists of three layers, namely a substrate, a cover plate and a metal supporting piece, wherein the substrate is provided with a microchannel for cell suspension to continuously flow and a microelectrode array for electrically stimulating cells; and the microelectrode array is engraved on the two sides of the microchannel, is connected with the metal supporting piece by a metal lead through a micropore, and is connected with a direct-current power supply through a wire to lead in an electrical signal. The microelectrode array chip has a simple structure and is provided with a small number of microelectrodes, so that processing difficulty is far lower than that of a high-density microelectrode cell electrofusion chip; the required fusion voltage is direct-current low voltage, the requirement on equipment is low, and device cost is far lower than that of the conventional cell electrofusion method; meanwhile, cell fusion is continuously performed in the process that cell pairs flow through the microchannel, fusion yield is also far higher than that of the conventional cell electrofusion method, so that the popularization and application of the microelectrode array chip are facilitated.
Description
Technical field
The present invention relates to biomass cells electricity integration technology.Be specifically related to a kind of chip apparatus that can under the low-voltage DC signal conditioning, realize successive cytogamy, comprise at the uniform velocity the flowing of sample introduction, cell, cell electroporation and the fusion of the design of microchannel and microelectrode, cell, the collection of fused cell.
Background technology
Cell fusion method comes from zooblast research.Japanese scholar deceived the Tian Shanxiong fusion of the Sendai virus of UV-light deactivation at external evoked ehrlich ascites tumor cell in 1962, had realized the artificial cell fusion method first.But the cytogamy productive rate of virus induction is extremely low, is difficult to satisfy growing research and application demand, and scientist begins to seek better to merge induction method.The high state of Canadian scientist of Chinese origin nanmu in 1974 finds that (polyethylene glycol PEG) can impel plant protoplast to merge to polyoxyethylene glycol, has opened up the cell fusion method of chemical induction, makes cell confluency than the virus induction method large increase arranged.Because cytogamy is in the huge applications potentiality of aspects such as biology, medical science, pharmacy, join in this research direction in succession from the various countries scientist in fields such as biology, medical science, physics.Last century the eighties, U.S. scientist Zimmermann is used for cytogamy with emerging electronic technology, take the lead in having proposed cell electricity fusion method, promptly under the effect of high frequency alternating electric field, make the cell polarization form electric dipole, and then along the power line direction bunchiness that sticks to each other, give the instantaneous high pressure pulse then, make the electrical breakdown of cytolemma reversibility, thereby cause cytogamy.Compare with the virus-mediated and chemical fusion method of tradition, the advantage that cell electricity merges is the fusion rate height, and is easy and simple to handle, quick, and the pair cell toxicological harmless can microscopic examination, induces the process controllability strong, is suitable for instrument application and standard operation etc.Therefore, cell electricity fusion method has just obtained widespread use after proposing.
Although the traditional electrical fusion method has obtained widespread use in cytogamy research, allos cell pairing remains at random to be carried out, and it is uncontrollable to merge direction, the higher but target of fusion rate join type to be fused into power still very low.And, the also difficult people's will to the greatest extent in aspects such as automation of operation, cost, productive rate, fused cell separation.From last century end to the beginning of this century, along with micro-processing technology and MEMS (micro electro mechanical system) (micro electro mechanical systems, MEMS) fast development of technology, the control of micro-operation method pair cell are more accurately effective, and the cell electricity merges research and also progresses into the microcosmic level.Just began in 1989 to realize that with micro fluidic device the cell electricity merges as people such as Japanese scientist Musuda.Recent years, microchip single cell electroporation and cell dielectric electrophoresis control techniques have obtained a lot of new achievements, a plurality of in the world laboratories combine these new technologies in succession with cell electricity integration technology, in the hope of solving the difficulty that conventional cell electricity fusion method is run into.
Aspect controllability, observability and the fusion rate large increase is being arranged although the application of microsystems technology is merged the cell electricity, also do not having the accurate pairing of fine solution allos cell, the quick acquisition of a large amount of fused cells and the several difficult such as effective separation of fused cell.Though utilize the micro-operation under the micro-condition can realize accurate pairing, it is a cost to sacrifice efficient.The people such as Skelley of U.S. MIT have proposed a kind of micro-current controlled cell pairing and fusion method recently, make the pairing rate of allos cell that large increase arranged.But the microchip structure that this method adopted is quite complicated, the processing request height, and the cell size is also restricted.And the same with existing most cells electricity fusion methods, cytogamy is batch to carry out, and promptly a large amount of cells enter chip simultaneously, carries out the separation and Extraction operation more simultaneously after finishing a mixing operation.Therefore, the fusion productive rate of whole device is still not high.In methods such as two-dimentional microelectrode array cytogamy, adopt a large amount of electrodes can improve the fusion amount, but owing to merge the restriction of cavity size, output is still very limited, is difficult to satisfy the research or the application demand of a large amount of fused cells of those needs.And number of electrodes is big, and difficulty of processing also significantly improves.The Continuous Flow operation is the effective means that overcomes the batch operation inefficiency in the microfluidic system, effectively realizes the Continuous Flow operation but can not find suitable structure design during cell electrofusion chip is studied in early days always.People such as Wang had once proposed a kind of successive direct current fusion method, utilized micro-processing technology to form a series of slits in a straight channel, loaded high voltage direct current at the electrode at passage two ends.Because the power line at slit place is crypto set more, strength of electric field is also higher, when cell to continuously by these slits the time, realize that under instantaneous high field intensity effect perforation merges.This method provides a kind of new thinking for realizing the continuous flow cell operation, is expected to solve problems such as cytogamy efficient is not high, emerging system complexity.But in this method, the use pair cell of high-voltage and large electrode activity and aseptic technique all bring disadvantageous effect.
Relevant patent is as follows both at home and abroad:
200610054121.x, 2006, University Of Chongqing, Zhao Zhiqiang etc.;
CN1482234,2003, Shanghai Inst. of Technical Physics, Chinese Academy of Sciences, Zhang Tao etc.;
CN86210174, nineteen ninety-five, institute of oncology, Liaoning, Liang Wei;
4326934，April?27，1982，Pohl；
441972，April?10，1982，Pohl；
4578168，March?25,?1986,?Hofman;
4695547,?September?22,?1987,?Hillard;
4699881,?October?13,?1987,?Matschke?et.?al;
5007995,?April?16,?1991,?Takahizuki。
Summary of the invention
The objective of the invention is at above-mentioned the deficiencies in the prior art, propose a kind of continuous flow cell electrofusion chip,, realize that under the small voltage condition cell perforation merges by the microelectrode array design based on microelectrode array and low dc voltage operation.Adopt low dc voltage electric field rather than high-voltage alternating electric field, lower to the requirement of the collecting of electrical controls, reduced the complicacy and the cost that merge instrument, the damage of pair cell is also corresponding to be reduced.Simultaneously, the operation of Continuous Flow can allow fusion process constantly carry out, and improves greatly to merge productive rate.
Technical scheme of the present invention is as follows:
A kind of microelectrode array chip that is used for the fusion of continuous flow cell electricity, described chip comprises substrate and cover plate, the microchannel of Gong the cell suspension continuous flow that has micro-processing method to scribe in the described substrate to form and give the microelectrode array of cell electricity irritation, cover plate bonding or be bonded in the substrate, turnover sample duct is arranged on it, be communicated with suprabasil microchannel.
The two bottom sides of described substrate respectively is fixed with the metallic support sheet.
Described substrate is made of siliceous stratum basale, silicon dioxide insulating layer, low-resistance silicon electrode layer and silicon dioxide protective film from bottom to up successively; adopt micro-processing technology etched recesses to silicon dioxide insulating layer on low-resistance silicon electrode layer 7 to form the microchannel; behind substrate and cover plate bonding; the microchannel forms the stream of sealing, and liquid can only flow in the microchannel.
Both sides at the region intermediate of the microchannel of described substrate, be to the microchannel medullary ray by the low-resistance silicon electrode layer that the broach shape is outstanding to form a series of microelectrodes, the microelectrode of both sides distributes in pairs, constitute microelectrode array, the microelectrode electrical communication of phase the same side is isolated by silicon dioxide insulating layer between the microelectrode of both sides.
At substrate on two sides edge, described microchannel; reserve the low-resistance silicon electrode layer that a part does not cover silicon dioxide protective film; form the lead-in wire stub-out; metal lead wire is respectively via the lead-in wire stub-out of both sides; be connected with the metallic support sheet of substrate two bottom sides; the metallic support sheet is incorporated into chip by the lead external direct current power supply with electrical signal.
In the cell fusion process, at first utilize biochemical method that two cell couplings are associated in together, use the micro-sampling pump to be advanced in chip by the sample introduction duct on the cover plate cell suspension then and make its at the uniform velocity continuous flow in the microchannel, the low dc voltage electrical signal that direct supply produces is loaded on the microelectrode array by plain conductor, be distributed in the electrical signal that loads opposed polarity on the microelectrode of both sides, microchannel respectively, small spacing forms local high field intensity electric field between electrode pair, be similar to and act on cell flowing through the right snap of cell between them last electricimpulse, thereby make membrane perforation and fusion, the cell of fusion successively flows out chip by sample outlet porous channel to be collected.
Base material of the present invention can adopt mechanically resistant materials such as silicon, glass, passes through micro-processing method etching one or more microchannel in substrate.The conductive substrates material after the microchannel machines, need treatment channel surface and with metallic support sheet adjacently situated surfaces, guarantee it can with microelectrode of processing later and the insulation of metallic support sheet.
Described microchannel is a beeline channel.The typical sizes of considering biomass cells is at 1 ~ 50 μ m, and the microchannel degree of depth in the chip and width change according to the difference of experimental cell, and channel depth is set in 40 ~ 100 μ m usually, and width is set in 100 ~ 200 μ m.
The microelectrode width is 20 ~ 200 μ m, and the distance of electrode end and medullary ray is decided according to microchannel and experimental cell size, generally at 20 ~ 50 μ m.Described microelectrode is and is distributed in the both sides, microchannel in pairs, microelectrode to number generally at 3 ~ 10 pairs.Microelectrode is divided into symmetric form at the relative arrangement mode of both sides, microchannel, and promptly the microelectrode on both sides presents the arrangement mode of horizontal symmetrical, perhaps alternative form, and promptly the microelectrode on both sides presents staggered relatively arrangement mode.The microelectrode of alternative form can be given and the cell electricity irritation in orthogonal direction distributing, and helps improving the fusion rate of cell.The microelectrode processing of phase the same side interconnects on same electrical conductor or with electro-conductive material, is distributed in the microelectrode mutual insulating of passage both sides.Microelectrode array is connected through the metallic support sheet of metal lead wire below substrate, links to each other with external source by lead then, and the microchannel between the microelectrode is the servicesubway of cytogamy.The metallic support sheet connects the lead-in wire from passage one side microelectrode respectively for separately two.
Microelectrode structure unanimity in the described microelectrode array is circular cylindrical shape, rectangle column, Polygons column or is circular taper, rectangle taper, Polygons cone structure
After substrate and cover plate lump together, the two ends, microchannel link to each other with sample outlet porous channel with the sample introduction duct, and external catheter inserts in the access opening fixing, make the inside and outside stream of chip be connected to form the stream of complete closed, can get rid of extraneous contamination, realize the asepticize operation of fusion process.Cell suspension can pump in the chip by the sample introduction duct, and the continuously and smoothly is flowed in the microchannel.To when the relative microelectrode, because the high strength of electric field in this zone, cell is subjected to the electricity irritation of similar electricimpulse to meeting at cell, and through after several identical stimulations, cell perforation takes place and merges.Cytogamy is successively carried out in flow process, can reach the purpose of continuous, a large amount of fused cells, the fusion productive rate of raising cell.
Continuous flow cell electrofusion chip based on microelectrode array can adopt micro-processing methods such as unicircuit (IC) complete processing, MEMS complete processing and soft lithographic complete processing.
The present invention has following technological merit compared with prior art:
1, continuous flow cell merges: cytogamy is successively carried out in the cell suspension flow process, can not be subjected to merging in the conventional cell fusion method restriction of pond volume, its productive rate, the level of automation of use and anti-pollution, anti-interference degree all improve a lot.
2, make microchannel and microelectrode with micro-processing method, shortened the spacing of comparative electrode, under the low voltage condition, can obtain the strength of electric field that cytogamy needs, guaranteed operator safety, also reduced cell injury.Simultaneously, utilize cell that the interelectrode electricimpulse effect of flowing through is made cytogamy, only need the DC voltage-stabilizing input to realize, needing in the traditional method to have avoided non-DC voltage-stabilizing signal input such as sinusoidal ac signal, pulse signal, the power supply signal equipment requirements is reduced greatly, better reliability, system cost is lower.
3, electrode pair quantity is 3 ~ 10 pairs in the chip, is less than thousands of right numbers in the conventional cell electrofusion chip greatly.Therefore, chip structure is simple, and difficulty of processing and cost all reduce greatly.
Description of drawings
Fig. 1 is based on the continuous flow cell electrofusion chip synoptic diagram of microelectrode array;
Fig. 2 is based on the continuous flow cell electrofusion chip structure wiring layout of microelectrode array;
Fig. 3 is based on the continuous flow cell electrofusion chip substrate synoptic diagram of microelectrode array;
Fig. 4 A is microelectrode array structural representation a---symmetric form;
Fig. 4 B is microelectrode array structural representation a---alternative form.
Embodiment
Referring to Fig. 1 and Fig. 2, Continuous Flow microelectrode array cell electrofusion chip is made up of turnover sample conduit 3, the metallic support sheet 4 of substrate 1, cover plate 2, the inside and outside stream of connection.
The structure of the substrate 1 of chip is referring to Fig. 3; adopt silicon materials; from bottom to up successively by siliceous stratum basale 5; silicon dioxide insulating layer 6; low-resistance silicon electrode layer 7 and silicon dioxide protective film 8 constitute; adopt micro-processing technology etched recesses to silicon dioxide insulating layer 6 on low-resistance silicon electrode layer 7 to form microchannel 9; the microchannel 9 on both sides is linear; the microchannel of region intermediate is distributed with a series of microelectrodes 10 of broach shape in the passage both sides; the microelectrode of these broach shapes is to be broach shape by low-resistance silicon electrode layer 7 to the microchannel centerline direction in the both sides of microchannel to give prominence to and form; microelectrode on the conduit wall of both sides distributes in pairs; constitute microelectrode array; isolate by the silicon dioxide insulating layer on surface between the microelectrode of both sides, and the microelectrode of the same side this be conducting.Behind substrate 1 and silex glass cover plate 2 bondings, the microchannel forms the stream of sealing, and liquid can only flow in passage.According to the difference of chip size, can integrated one on chip to many microchannels, the yardstick of electrode and interelectrode spacing can be adjusted according to the difference of subjects, the electrode pair number is 3 ~ 10 pairs.At substrate on two sides edge, microchannel; reserve the low-resistance silicon electrode layer that a part does not cover silicon dioxide protective film; form lead-in wire stub-out 11; metal lead wire 12 is respectively through adopting bonding or weldprocedure to be drawn by the lead-in wire stub-out 11 of both sides; link to each other with the metallic support sheet 4 of a side separately, metallic support sheet 4 passes through wire connecting power.The linear pattern microchannel of constructing on this chip helps flowing of cell suspending liquid, reduces cell sticking in flow process as far as possible, guarantees mobile stability and reduces the cell alluvial.Behind the chip bonding, in the injection port 13 of cover plate and outlet 14, insert turnover sample conduit 3 respectively.In the fusion experiment, utilize biochemical coupling connection (after the vitamin H cell marking, with avidin the cell of mark is connected together) cell in the cell suspension is connected together to form cell right, cell suspension enters chip by turnover sample conduit 3 and injection port 13 under the driving of micro pump then, cell suspension is uniform motion in microchannel 9, when cell when flowing through a pair of microelectrode 10, microelectrode to the high field intensity electric field with pair cell to applying electricity irritation, because cell is ofer short duration to the right time of the microelectrode of flowing through, this stimulates the electricimpulse in the similar conventional cell fusion method, and pulse operating time is the time of cell to this electrode pair of flowing through.Perforation takes place and merges in cell under a series of electricity irritation effects, fused cell flows into collection container through outlet 14 and conduit 3.Microchannel 9, injection port 13, outlet 14, the conduit 3 of sealing constitutes the stream of sealing together, can carry out continuously, operate automatically by the pair cell suspension, improves operation efficiency and reliability, can also get rid of external interference.Cytogamy operation continuously also can obtain a large amount of fused cells at short notice, improves productive rate, reaches the purpose of efficient cytogamy.
Fig. 4 shows is the array formed of microelectrode 10 and the part of microchannel 9, microelectrode 10 is distributed in the both sides of microchannel 9, the microelectrode arrangement mode is divided into symmetric form (seeing Fig. 4 A) on the microelectrode array relatively, it is the arrangement mode that microelectrode presents horizontal symmetrical, perhaps alternative form (seeing Fig. 4 B), promptly microelectrode presents staggered symmetric arrangement mode and arranges.The degree of depth of microchannel 9 is 50 μ m, and width is 140 μ m, and the end-to-end distance of microelectrode 10 is 20 μ m from channel centerline.The diameter of injection port 13 and outlet 14 is 2500 μ m, utilizes laser cutting method processing.Turnover sample conduit 3 is of a size of internal diameter 800 μ m, and external diameter 2500 μ m are with abundant assurance cell admission passage inside smoothly.
Among Fig. 3, siliceous stratum basale 5 is of a size of 2 cm * 3 cm, and thickness is 500 μ m, and the support of sufficient intensity is provided for chip; Silicon dioxide insulating layer 6 is of a size of 2 cm * 3 cm, and thickness is 10 μ m, makes chip not be afraid of high-intensity electrostatic breakdown, ensures the electric stability of chip; Low-resistance silicon electrode layer 7 is of a size of 2 cm * 3 cm, and thickness is 50 μ m, promptly merges the degree of depth of passage.Very little, the good mechanical property of low-resistance silicon internal stress, low-resistivity can improve the chip electric property, improves fusion efficiencies.On low-resistance silicon electrode layer 7, realize silicon dioxide protective film 8, improve the resistance of oxidation of chip by plasma-reinforced chemical vapor deposition method (PECVD) technology.Metallic support sheet 4 is of a size of 0.5 cm * 3 cm, and thickness is 0.5 mm.
The complete processing of above-mentioned continuous flow cell electrofusion chip is:
(1) chooses top layer silicon the thick silicon on insulator material of 50 μ m is arranged;
(2) isolation diffusion is finished top layer low-resistance silicon degree of depth knot;
(3) top layer low-resistance silicon oxidation, oxidated layer thickness are 550 ~ 600 nm;
(4) photoetching silicon-dioxide;
(5) sputter sial Si-Al forms trace layer, and trace layer thickness is 2 ± 0.2 μ m;
(6) photoetching lead-in wire;
(7) alloy;
(8) plasma-reinforced chemical gaseous phase deposition pecvd process is realized the silicon dioxide insulator passivation, and silicon dioxide passivation layer thickness is 1.5 ± 0.2 μ m;
(9) photoetching;
(10) dry etching silicon-dioxide;
(11) dry etching silicon, the degree of depth are 50 μ m, until insulation layer, do not link to each other mutually on each interdigitated electrodes electrical structure to allow, and form the microchannel;
(12) dry method is removed photoresist;
Microelectrode array substrate that utilization machines and cover plate bonding, chip that bonding is good and metallic support sheet are fixed by tamanori, each microelectrode array on the chip and corresponding metallic support sheet are linked to each other with spun gold by bonding techniques then, conduit is inserted injection port and outlet forms airtight microchannel.
Claims (5)
1. one kind is used for the microelectrode array chip that the continuous flow cell electricity merges, described chip comprises substrate and cover plate, the microchannel of Gong the cell suspension continuous flow that has micro-processing method to scribe in the described substrate to form and give the microelectrode array of cell electricity irritation, cover plate bonding or be bonded in the substrate, turnover sample duct is arranged on it, be communicated with suprabasil microchannel; It is characterized in that:
The two bottom sides of described substrate respectively is fixed with a metallic support sheet;
Described substrate is made of siliceous stratum basale, silicon dioxide insulating layer, low-resistance silicon electrode layer and silicon dioxide protective film from bottom to up successively, adopt micro-processing technology etched recesses to silicon dioxide insulating layer on the low-resistance silicon electrode layer to form the microchannel, behind substrate and cover plate bonding, the microchannel forms the stream of sealing, and liquid can only flow in the microchannel;
Both sides at the region intermediate of the microchannel of described substrate, be to the microchannel medullary ray by the low-resistance silicon electrode layer that the broach shape is outstanding to form a series of microelectrodes, the microelectrode of both sides distributes in pairs, constitute microelectrode array, the microelectrode electrical communication of phase the same side is isolated by silicon dioxide insulating layer between the microelectrode of both sides;
At substrate on two sides edge, described microchannel, reserve the low-resistance silicon electrode layer that a part does not cover silicon dioxide protective film, form the lead-in wire stub-out, metal lead wire is respectively via the lead-in wire stub-out of both sides, be connected with the metallic support sheet of substrate two bottom sides, the metallic support sheet is incorporated into chip by the lead external direct current power supply with electrical signal;
In the cell fusion process, at first utilize biochemical method that two cell couplings are associated in together, use the micro-sampling pump to be advanced in chip by the sample introduction duct on the cover plate cell suspension then and make its at the uniform velocity continuous flow in the microchannel, the low dc voltage electrical signal that direct supply produces is loaded on the microelectrode array by plain conductor, be distributed in the electrical signal that loads opposed polarity on the microelectrode of both sides, microchannel respectively, small spacing forms local high field intensity electric field between electrode pair, be similar to and act on cell flowing through the right snap of cell between them last electricimpulse, thereby make membrane perforation and fusion, the cell of fusion successively flows out chip by sample outlet porous channel to be collected.
2. the microelectrode array chip that is used for the fusion of continuous flow cell electricity according to claim 1, it is characterized in that: described microchannel is a beeline channel, and the microchannel degree of depth is set in 40 ~ 100 μ m, and width is set in 100 ~ 200 μ m.
3. the microelectrode array chip that is used for the fusion of continuous flow cell electricity according to claim 1 and 2, it is characterized in that: the width of described microelectrode is 20 ~ 200 μ m, the terminal distance with the microchannel medullary ray of microelectrode is 20 ~ 50 μ m.
4. according to claim 1, the 2 or 3 described microelectrode array chips that are used for the fusion of continuous flow cell electricity, it is characterized in that: the arrangement mode that the microelectrode of both sides, described microchannel is relative is symmetric form, perhaps alternative form; The right number of microelectrode is 3 ~ 10 pairs.
5. according to claim 1, the 2 or 3 described microelectrode array chips that are used for the fusion of cell electricity, it is characterized in that: the microelectrode structure unanimity in the described microelectrode array is circular cylindrical shape, rectangle column, Polygons column or is circular taper, rectangle taper, Polygons cone structure.
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102517207A (en) * | 2011-12-02 | 2012-06-27 | 重庆大学 | Cellular electric fusion chip device based on dispersed type side wall microelectrode array and processing process |
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| CN103074212A (en) * | 2013-01-23 | 2013-05-01 | 苏州大学 | Cell electrofusion device and method |
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| 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 |
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101168724A (en) * | 2007-10-26 | 2008-04-30 | 重庆大学 | Cell electrofusion chip and processing method thereof |
| CN101250482A (en) * | 2008-03-28 | 2008-08-27 | 重庆大学 | Microelectrode array chip for cell electrofusion |
| CN101343613A (en) * | 2008-08-22 | 2009-01-14 | 重庆大学 | Flexible high-pass cell electric amalgamation microelectrode array chip apparatus |
| CN101368155A (en) * | 2008-09-19 | 2009-02-18 | 重庆大学 | Continuous flow cell electric amalgamation chip based on silicon structure on insulator |
| CN101693874A (en) * | 2009-09-30 | 2010-04-14 | 重庆大学 | Cell electrofusion chip device based on micro-chamber array structure |
| CN101693875A (en) * | 2009-09-30 | 2010-04-14 | 重庆大学 | Cell electrofusion chip device based on columnar microelectrode array and electrofusion method |
-
2011
- 2011-01-25 CN CN2011100269211A patent/CN102174387A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101168724A (en) * | 2007-10-26 | 2008-04-30 | 重庆大学 | Cell electrofusion chip and processing method thereof |
| CN101250482A (en) * | 2008-03-28 | 2008-08-27 | 重庆大学 | Microelectrode array chip for cell electrofusion |
| CN101343613A (en) * | 2008-08-22 | 2009-01-14 | 重庆大学 | Flexible high-pass cell electric amalgamation microelectrode array chip apparatus |
| CN101368155A (en) * | 2008-09-19 | 2009-02-18 | 重庆大学 | Continuous flow cell electric amalgamation chip based on silicon structure on insulator |
| CN101693874A (en) * | 2009-09-30 | 2010-04-14 | 重庆大学 | Cell electrofusion chip device based on micro-chamber array structure |
| CN101693875A (en) * | 2009-09-30 | 2010-04-14 | 重庆大学 | Cell electrofusion chip device based on columnar microelectrode array and electrofusion method |
Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102517207B (en) * | 2011-12-02 | 2013-12-18 | 重庆大学 | Cellular electric fusion chip device based on dispersed type side wall microelectrode array and processing process |
| CN102517207A (en) * | 2011-12-02 | 2012-06-27 | 重庆大学 | Cellular electric fusion chip device based on dispersed type side wall microelectrode array and processing process |
| CN103013821A (en) * | 2012-12-03 | 2013-04-03 | 清华大学 | Avidin-biotin system cell patterning-based chip and preparation and applications thereof |
| CN103074212A (en) * | 2013-01-23 | 2013-05-01 | 苏州大学 | Cell electrofusion device and method |
| CN103451097B (en) * | 2013-08-19 | 2015-05-27 | 西安交通大学 | In-vitro cell fusion system and method for increasing bone marrow transplantation success rate |
| CN103451097A (en) * | 2013-08-19 | 2013-12-18 | 西安交通大学 | In-vitro cell fusion system and method for increasing bone marrow transplantation success rate |
| CN104531524A (en) * | 2014-12-11 | 2015-04-22 | 国家纳米科学中心 | Micro-needle tip array chip for cell electroporation and application of micro-needle tip array chip |
| 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 |
| CN106676001A (en) * | 2016-12-29 | 2017-05-17 | 中国科学院微电子研究所 | Cell electroporation system and method based on selective continuous flow of crossed narrow channels |
| CN106676001B (en) * | 2016-12-29 | 2019-07-30 | 中国科学院微电子研究所 | Cell electroporation system and method based on selective continuous flow of crossed narrow channels |
| WO2019076353A1 (en) * | 2017-10-19 | 2019-04-25 | 苏州壹达生物科技有限公司 | Flow electroporation device |
| CN108330067A (en) * | 2018-05-14 | 2018-07-27 | 上海启文生物科技有限公司 | Integrated casket shape cell fusion electrode |
| CN108330067B (en) * | 2018-05-14 | 2023-08-25 | 上海启文生物科技有限公司 | Integrated casket-like cell fusion electrode |
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