CN108795708A - A kind of micro-fluidic chip carrying out cell aggregation based on microelectrode array - Google Patents
A kind of micro-fluidic chip carrying out cell aggregation based on microelectrode array Download PDFInfo
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- CN108795708A CN108795708A CN201811091200.7A CN201811091200A CN108795708A CN 108795708 A CN108795708 A CN 108795708A CN 201811091200 A CN201811091200 A CN 201811091200A CN 108795708 A CN108795708 A CN 108795708A
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- microelectrode
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS 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
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/02—Form or structure of the vessel
- C12M23/16—Microfluidic devices; Capillary tubes
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS 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
- C12M35/00—Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
- C12M35/02—Electrical or electromagnetic means, e.g. for electroporation or for cell fusion
Abstract
A kind of micro-fluidic chip carrying out cell aggregation based on microelectrode array, the technical issues of being related to microfluidic art, collected the solution is to high-flux cell.The chip includes the container floor stacked gradually from top to bottom, microelectrode layer, basal layer；The container floor is equipped at least one container hole and at least two electrode holes, and positive electricity pole pipe, negative electrode tube are plugged with respectively in each electrode hole；The microelectrode layer is made of at least one planar electrode array, the microelectrode piece that each planar electrode array is tiled by even number forms, the hole heart that each microelectrode piece of each planar electrode array surrounds corresponding container hole is symmetrically laid, and each microelectrode piece is divided into positive microelectrode piece, negative microelectrode piece, each positive microelectrode piece is coupled with each positive electricity pole pipe, and each negative microelectrode piece is coupled with each negative electrode tube.Chip provided by the invention may be implemented high-throughput various kinds of cell and collect.
Description
Technical field
The present invention relates to microflow control techniques, and the micro-fluidic of cell aggregation is carried out based on microelectrode array more particularly to a kind of
The technology of chip.
Background technology
Microfluidic chip technology is to be put forward for the first time by the Manz and Widmer of Switzerland early 1990s, due to its tool
Have analyze speed is fast, reagent consumption less, be easily integrated and many advantages, such as high throughput analysis, over the last couple of decades, miniflow
Control chip achieves significant progress in terms of the analyzing processing of biological and chemical sample, in recent years microfluidic chip technology
Researcher has been transferred to research direction increasingly complex biosystem, such as carries out cell analysis using micro-fluidic chip
The research such micro-fluidic chips of play more the exploratory development in the fields such as cell biology, neurology, pharmacy and organizational project
Carry out more important role.
Micro-fluidic chip cell operation is that cell is transported to scheduled site, after being fixed, carries out cell component, knot
The analyses such as structure and function.Existing microflow control technique all makes micro- well, micropore, Wei Ba, micro- slit and micro-pipe using micro Process mode
The geometry trap such as road or obstacle capture cell, can not only form open array system, can also be realized to thin in microchannel
The control of born of the same parents.But the processing request of micro Process mode is higher, and microchannel is easy blocking, preparation of samples requires high, discomfort
Close the collection of high-throughput various kinds of cell.
Invention content
Defect present in for the above-mentioned prior art, technical problem to be solved by the invention is to provide one kind to be not easy to block up
Plug, it is low to sample requirement, and the micro-fluidic chip that cell aggregation is carried out based on microelectrode array of handling ease.
In order to solve the above-mentioned technical problem, provided by the present invention a kind of based on the micro- of microelectrode array progress cell aggregation
Fluidic chip, it is characterised in that：Including the container floor, microelectrode layer, basal layer stacked gradually from top to bottom；
The container floor is equipped at least one container hole and at least two electrode holes, and the container hole, electrode hole open upwards,
Conductive electrode tube there are one being planted in each electrode hole；
Each electrode tube is divided into two groups, and one group of electrode tube therein is positive electricity pole pipe, and another group of electrode tube is negative electrode tube；
The microelectrode layer is made of at least one planar electrode array, micro- electricity that each planar electrode array is tiled by even number
Pole piece forms, and each planar electrode array of microelectrode layer and each container hole of container floor correspond；
The hole heart that each microelectrode piece of each planar electrode array surrounds corresponding container hole is symmetrically laid, and each micro- electricity
Pole piece is divided into two groups, and one group of microelectrode piece therein is positive microelectrode piece, and another group of microelectrode piece is negative microelectrode piece, each
Positive microelectrode piece is coupled with each positive electricity pole pipe, and each negative microelectrode piece is coupled with each negative electrode tube.
Further, each microelectrode piece of each planar electrode array has the lower end of regional area and corresponding container hole
Aperture is overlapped.
Further, the microelectrode piece is gold electrode piece.
Further, the electrode tube is the copper pipe of surface gold-plating.
Further, the basal layer is the slide that thickness is less than 2 millimeters.
The micro-fluidic chip provided by the invention that cell aggregation is carried out based on microelectrode array, using flat geometry structure
Type makes microelectrode piece on the base layer, applies the control voltage that voltage signal is collected as dielectrophoresis in microelectrode on piece,
To make dielectrophoresis process be realized on each container hole, container hole, electrode hole aperture can design it is relatively large, because
This is relatively low to the processing request of container hole, electrode hole, and container hole is not easy to block, and the requirement to sample is relatively low.
Description of the drawings
Fig. 1 is that the plan structure of the micro-fluidic chip for carrying out cell aggregation based on microelectrode array of the embodiment of the present invention is shown
It is intended to；
Fig. 2 is the left view cut away view of the micro-fluidic chip that cell aggregation is carried out based on microelectrode array of the embodiment of the present invention.
Specific implementation mode
The embodiment of the present invention is described in further detail below in conjunction with description of the drawings, but the present embodiment is not used to limit
The system present invention, every similar structure using the present invention and its similar variation, should all be included in protection scope of the present invention, the present invention
In pause mark indicate that the relationship of sum, the English alphabet in the present invention are case sensitive.
As Figure 1-Figure 2, what the embodiment of the present invention was provided is a kind of based on the micro- of microelectrode array progress cell aggregation
Fluidic chip, it is characterised in that：Including the container floor 1, microelectrode layer 2, basal layer 3 stacked gradually from top to bottom；
The container floor 1 is equipped with multiple containers hole 11 and multiple electrodes hole 12, and the container hole 11, electrode hole 12 are opened upwards
It puts, there are one conductive electrode tubes 13 for plant in each electrode hole 12；
Each electrode tube 13 is divided into two groups, and one group of electrode tube 13 therein is positive electricity pole pipe, and another group of electrode tube 13 is negative electrode
Pipe；
The microelectrode layer 2 is made of multiple planar electrode arrays, the microelectrode piece that each planar electrode array is tiled by four
21 compositions, each planar electrode array of microelectrode layer 2 are corresponded with each container hole 11 of container floor 1；
The hole heart that four microelectrode pieces 21 of each planar electrode array surround corresponding container hole 11 is symmetrically laid, and each
Microelectrode piece 21 has that regional area is Chong Die with the lower end aperture of container hole 11, and four microelectrode pieces 21 are divided into two groups,
One group of microelectrode piece 21 therein is positive microelectrode piece, and another group of microelectrode piece 21 is negative microelectrode piece, each positive microelectrode piece
21 are coupled with each positive electricity pole pipe 13, and each negative microelectrode piece 21 is coupled with each negative electrode tube 13.
In the embodiment of the present invention, the container hole number on the container floor has 1 to 200, and the preferred aperture of container hole is 4
Millimeter, there are four the microelectrode pieces in planar electrode array, the gold electrode piece of 100 nanometers of microelectrode piece preferred thickness, same flat
The laying spacing of each microelectrode piece in the electrod-array of face is 100um；The copper pipe of the preferred surface gold-plating of electrode tube in electrode hole；
In other embodiments of the present invention, the quantity of the microelectrode piece in planar electrode array can also be other quantity, but microelectrode
The quantity of piece should be even number（Such as 2,6 etc.）, the quantity of positive and negative microelectrode piece can be made consistent in this way, electrode tube
The pipe fitting, such as pure bobbin etc. that can be conductive of other materials can also be used.
In the embodiment of the present invention, the basal layer is that the thickness that is prepared by glass, quartz or calcirm-fluoride is saturating less than 2 millimeters
Bright slide, the microelectrode piece in planar electrode array on the base layer, then by bonding mode connect container by coated film deposition
Layer.
The voltage signal generator that cooperation of the embodiment of the present invention can export 0.5V to 50V voltages uses, and high pass may be implemented
The various kinds of cell of amount is collected, stimulation of the parallel parsing different pharmaceutical concentration to different cells and its real-time observation in piece and
Detection can be used for various concentration concurrent effect various kinds of cell, carry out high-flux cell drug susceptibility screening.
For collecting Escherichia coli, Escherichia coli solution is injected into each container hole, by the electricity of voltage signal generator
Pressure signal output end is connected to the electrode tube in each electrode hole, and voltage signal generator exports the voltage signal of an alternate peaks
（Such as the 4V alternating currents that frequency is 1MHz）The electrode tube being loaded into each electrode hole, and then be loaded into planar electrode array
Each microelectrode piece, voltage signal driving under generate inhomogeneous field formed dielectrophoretic motion, resulting electrophoretic force
It is directed toward the center of planar electrode array（It is also the hole heart of container hole）, after a period of time（Such as 20 minutes）, aobvious by fluorescence
Micro mirror can see the Escherichia coli being collected into and form a compact three-dimensional globoid.
Dielectrophoresis is the lower object of dielectric constant orientation of stress generation due to polarization in non-uniform electric field
Movement, can effectively realize being accurately positioned, detach, collect, capture and transporting for micron ion.Dielectric power size is with object
No electrification is unrelated, field strength, field strength change with the size, electrical properties, the electrical properties of surrounding medium and extra electric field of object
Rate, frequency are related.
Claims (5)
1. a kind of micro-fluidic chip carrying out cell aggregation based on microelectrode array, it is characterised in that：Including from top to bottom successively
Container floor, microelectrode layer, the basal layer of stacking；
The container floor is equipped at least one container hole and at least two electrode holes, and the container hole, electrode hole open upwards,
Conductive electrode tube there are one being planted in each electrode hole；
Each electrode tube is divided into two groups, and one group of electrode tube therein is positive electricity pole pipe, and another group of electrode tube is negative electrode tube；
The microelectrode layer is made of at least one planar electrode array, micro- electricity that each planar electrode array is tiled by even number
Pole piece forms, and each planar electrode array of microelectrode layer and each container hole of container floor correspond；
The hole heart that each microelectrode piece of each planar electrode array surrounds corresponding container hole is symmetrically laid, and each micro- electricity
Pole piece is divided into two groups, and one group of microelectrode piece therein is positive microelectrode piece, and another group of microelectrode piece is negative microelectrode piece, each
Positive microelectrode piece is coupled with each positive electricity pole pipe, and each negative microelectrode piece is coupled with each negative electrode tube.
2. the micro-fluidic chip according to claim 1 for carrying out cell aggregation based on microelectrode array, it is characterised in that：Often
Each microelectrode piece of a planar electrode array has regional area to be overlapped with the lower end aperture of corresponding container hole.
3. the micro-fluidic chip according to claim 1 for carrying out cell aggregation based on microelectrode array, it is characterised in that：Institute
It is gold electrode piece to state microelectrode piece.
4. the micro-fluidic chip according to claim 1 for carrying out cell aggregation based on microelectrode array, it is characterised in that：Institute
State the copper pipe that electrode tube is surface gold-plating.
5. the micro-fluidic chip according to claim 1 for carrying out cell aggregation based on microelectrode array, it is characterised in that：Institute
It is the slide that thickness is less than 2 millimeters to state basal layer.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811091200.7A CN108795708A (en) | 2018-09-19 | 2018-09-19 | A kind of micro-fluidic chip carrying out cell aggregation based on microelectrode array |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811091200.7A CN108795708A (en) | 2018-09-19 | 2018-09-19 | A kind of micro-fluidic chip carrying out cell aggregation based on microelectrode array |
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| CN108795708A true CN108795708A (en) | 2018-11-13 |
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| CN201811091200.7A Withdrawn CN108795708A (en) | 2018-09-19 | 2018-09-19 | A kind of micro-fluidic chip carrying out cell aggregation based on microelectrode array |
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Citations (7)
| 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 |
| JP2009287934A (en) * | 2008-05-27 | 2009-12-10 | Ulvac Japan Ltd | Microelectrode array device, its manufacturing method and bioassay method |
| JP2009288080A (en) * | 2008-05-29 | 2009-12-10 | Nippon Telegr & Teleph Corp <Ntt> | Extracellular micro electrode and method for manufacturing the same |
| CN102671724A (en) * | 2011-02-17 | 2012-09-19 | 王崇智 | Microelectrode array architecture |
| CN104789443A (en) * | 2014-06-23 | 2015-07-22 | 中国科学院微电子研究所 | Cellular localization unit, array, and device and formation method of cellular localization unit, array, and device |
| CN105219642A (en) * | 2015-09-09 | 2016-01-06 | 清华大学 | Catch and the micro-fluidic device of three-dimensional rotation for unicellular |
| CN106783851A (en) * | 2017-01-19 | 2017-05-31 | 北京世纪金光半导体有限公司 | SiCJFET devices of integrated schottky diode and preparation method thereof |
-
2018
- 2018-09-19 CN CN201811091200.7A patent/CN108795708A/en not_active Withdrawn
Patent Citations (7)
| 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 |
| JP2009287934A (en) * | 2008-05-27 | 2009-12-10 | Ulvac Japan Ltd | Microelectrode array device, its manufacturing method and bioassay method |
| JP2009288080A (en) * | 2008-05-29 | 2009-12-10 | Nippon Telegr & Teleph Corp <Ntt> | Extracellular micro electrode and method for manufacturing the same |
| CN102671724A (en) * | 2011-02-17 | 2012-09-19 | 王崇智 | Microelectrode array architecture |
| CN104789443A (en) * | 2014-06-23 | 2015-07-22 | 中国科学院微电子研究所 | Cellular localization unit, array, and device and formation method of cellular localization unit, array, and device |
| CN105219642A (en) * | 2015-09-09 | 2016-01-06 | 清华大学 | Catch and the micro-fluidic device of three-dimensional rotation for unicellular |
| CN106783851A (en) * | 2017-01-19 | 2017-05-31 | 北京世纪金光半导体有限公司 | SiCJFET devices of integrated schottky diode and preparation method thereof |
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Effective date of registration: 20210223 Address after: 519031 room 105-52680, No.6 Baohua Road, Hengqin New District, Zhuhai City, Guangdong Province (centralized office area) Applicant after: ZHUHAI HEJING TECHNOLOGY DEVELOPMENT Co.,Ltd. Address before: Room 227, building 7, 365 Chuanhong Road, Pudong New Area, Shanghai, 201202 Applicant before: SHANGHAI YINBO BIOLOGICAL TECHNOLOGY Co.,Ltd. |
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Effective date of registration: 20220928 Address after: Room 4292, building 24, No.2 Xincheng Road, Pudong New Area, Shanghai, 201306 Applicant after: SHANGHAI LELEE LASER TECHNOLOGY Co.,Ltd. Address before: 519031 room 105-52680, No.6 Baohua Road, Hengqin New District, Zhuhai City, Guangdong Province (centralized office area) Applicant before: ZHUHAI HEJING TECHNOLOGY DEVELOPMENT Co.,Ltd. |
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Application publication date: 20181113 |
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