CN106244416A - Cell microinjection chip based on dielectrophoresis, injection device and method of work thereof - Google Patents
Cell microinjection chip based on dielectrophoresis, injection device and method of work thereof Download PDFInfo
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- CN106244416A CN106244416A CN201610615682.6A CN201610615682A CN106244416A CN 106244416 A CN106244416 A CN 106244416A CN 201610615682 A CN201610615682 A CN 201610615682A CN 106244416 A CN106244416 A CN 106244416A
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- cell
- fluid channel
- microelectrode array
- microinjection
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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
- C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
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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
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/87—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
- C12N15/89—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation using microinjection
Abstract
The present invention relates to a kind of cell microinjection chip based on dielectrophoresis, injection device and method of work thereof, this cell microinjection chip includes: injection fluid channel and multistage fluid channel, wherein said multistage fluid channel is suitable to screen cell, so that only one cell enters in injection fluid channel, carry out microinjection；Cell microinjection chip, injection device and the method for work thereof of the present invention, to use corresponding microelectrode array to produce corresponding dielectrophoretic force, such as Conventional dielectric swimming power, row ripple dielectrophoretic force and rotation dielectrophoretic force, with control respectively signaling, rise or fall, pose adjustment, improving tradition microinjection chip utilizes micro syringe pump to drive the mode of its interior micro-fluid movement, reduction equipment complexity and cost, this cell microinjection chip can improve the efficiency of microinjection, it is simple to popularization and application.
Description
Technical field
The invention belongs to RESEARCH ON CELL-BIOLOGY technical field, a kind of experiment for cell microinjection fills
Put.
Background technology
Microinjection technique is based on intracellular microinjection and Micro-perfusion in Graft After technology, in increasing experimental biology
Research field becomes a microoperation technology the most universal, the most in vitro in fertilization, transgenic etc..These operations are
By single or multiple tubular glass micropipette, accurate positioner (micromanipulator) and microsyringe
Or microperfusion device is carried out on single cell.Microinjection is to utilize the glass of tip superfine (0.1 to 5 μm) micro-
Amount entry needle, is injected directly into exogenous genetic fragment in the cell of protokaryon phase embryo or cultivation, then by host genome sequence
Arrange contingent restructuring, lack, replicate or the phenomenon such as transposition and make exogenous gene embed in the chromosome of host.This micro-
The program of injection art, need to have quite accurate micromanipulator, when manufacturing long tip, need to elongate device with micropipet, injection
Time need to have the micromanipulation device of fixing tip position.The strong point of this technology is that any DNA the most all can be incoming any kind
Class intracellular.Compared with other transgenic technologys, the advantage of microinjection is that transgene efficiency is stable, injects gene size not
Restricted, without carrier and operation object without particular/special requirement etc., animals on the brink of extinction protection, reproduction and healthy, species improvement and
The research of intracellular caryoplasm is used widely.This method Successful utilization is in including that mice, fish, rat, rabbit and many are large-scale
Domestic animal, such as Transgenic animals such as cattle, sheep, pigs.
Micropipette capillary tube used in these operations draws pin device to make, and is first added by capillary glass tube
The temperature that heat melts to it, then it is drawn into diameter and the taper of required suitable size, the diameter of micropipette microcephaly
(little to 0.2 micron) is relevant to the high accuracy of fiber manipulator, and it may be used for moving accurately liquid.This degree of accuracy can be each
The cell of type and arbitrary size provide in the range of sub-picoliters level or 0.1 micron (micron) accurate, repeated well
Intracellular and cell sidenote penetrate.By using direct hydraulic pressure (pressure injection) or not by using current, and by executing
Add an electric field to make charged ion move (iontherapy), can obtain and the material in micropipette squeezed ejection go
Effect.
Its shortcoming is that equipment is accurate and expensive, operating technology needs exercise for a long time, and can only inject limited every time
Cell.And, cell, before cell is carried out microinjection, is not adjusted to by conventional cell microinjection device
Being suitable for " attitude " of injection, this will be substantially reduced the survival rate of cell after microinjection.Although presently, there are adjustment cell appearance
The device of state, but its by cell pose adjustment well after, microinjection device and adjust the device of cell attitude have can during injection
Can interfere or inharmonious and affect microinjection effect.It is necessary to propose further solution party for problem above
Case.
Summary of the invention
It is an object of the invention to provide a kind of cell microinjection chip, injection device and method of work thereof, with by being situated between
Electrophoretic force driving signaling is to injecting fluid channel, to complete injection.
In order to solve above-mentioned technical problem, the invention provides a kind of cell microinjection chip, including: injection fluid channel
With multistage fluid channel, wherein said multistage fluid channel is suitable to screen cell, so that only one cell enters injection miniflow
In road, carry out microinjection.
Further, described multistage fluid channel includes: at least two-stage fluid channel；I.e. one-level fluid channel and two grades of fluid channel, and
The width of flow path of next stage fluid channel is less than the width of flow path of upper level fluid channel；And the bottom of described one-level fluid channel is provided with
First microelectrode array, this first microelectrode array is suitable to drive cell suspension after powered up and move to two grades of fluid channel；Institute
The bottom stating two grades of fluid channel is provided with the second microelectrode array；When cell is after one-level fluid channel enters two grades of fluid channel, pass through
Second microelectrode array adjusts cell and enters hoverheight and the translational speed of injection fluid channel.
Further, the bottom of described injection fluid channel is provided with the 3rd microelectrode array, and top is provided with the 4th microelectrode array,
And the 4th the center of microelectrode array be provided with injection needle；Described 3rd microelectrode array is suitable to control cell to micro-
The underface of entry needle is moved, and by described 4th microelectrode array capture cell so that it is just it is being positioned injection needle
Lower section；And by, after the three, the 4th microelectrode array synergic adjustment cell attitudes, making cell move up or down；Carefully
When moving on born of the same parents, described microinjection is needled into cell, injects；When cell moves down, inject cell detachment microinjection
Pin, and entered output fluid channel by the 3rd microelectrode array by injecting cell.
Further, the bottom of described output fluid channel is provided with the 5th microelectrode array；And be energized at the 5th microelectrode array
After, make to inject cell output.
Further, described upper level fluid channel is horn mouth transition with the junction of lower two grades of fluid channel.
Further, described injection fluid channel, multistage fluid channel and output fluid channel are positioned in miniflow channel layer, and
The upper and lower quartz substrate that has been covered each by of described miniflow channel layer, described first, second, third and the 5th microelectrode
Array distribution is distributed in upper strata quartz substrate in lower floor's quartz substrate, the 4th microelectrode array.
Further, described first, the 3rd and the 5th microelectrode array is arranged at equal intervals by parallel pole and forms, and parallel
Electrode is orthogonal with the moving direction of cell；Described second microelectrode array uses two fork-shaped electrode fingers to form；And described
In four microelectrode arrays, each electrode is at the spaced 90 ° of angles of circumferencial direction, and extends to the central axis of injection needle.
Another aspect, present invention also offers a kind of cell microinjection device.
Described cell microinjection device includes: for connecting the control of each microelectrode array in cell microinjection chip
Module；Described control module produce each microelectrode array corresponding control signal, with control cell from multistage fluid channel move to
Injection fluid channel is to inject cell location, pose adjustment, and will to inject cell from entry needle after injection completes
Export after disengaging.
The third aspect, present invention also offers the method for work of a kind of cell microinjection device.
The method of work of described cell microinjection device comprises the steps:
Step S1, screens cell, makes only one cell enter in injection fluid channel；Step S2, micro-to entering to inject
Cell in runner is injected；And step S3, will inject cell output.
Further, the cell microinjection chip used in described cell microinjection device be suitable for use with described carefully
Born of the same parents' microinjection chip.
The invention has the beneficial effects as follows, cell microinjection chip, injection device and the method for work thereof of the present invention, to adopt
Corresponding dielectrophoretic force, such as Conventional dielectric swimming power, row ripple dielectrophoretic force and rotation dielectrophoretic force is produced with corresponding microelectrode array,
With control respectively signaling, rise or fall, pose adjustment, improve tradition microinjection chip utilize micro syringe pump to drive
The mode of its interior micro-fluid movement, reduces equipment complexity and cost, and this cell microinjection chip can improve micro-
The efficiency of injection, it is simple to popularization and application.
Accompanying drawing explanation
The present invention is further described with embodiment below in conjunction with the accompanying drawings.
Fig. 1 is the overall structure schematic diagram of the microinjection chip of the present invention；
Fig. 2 be the present invention miniflow channel layer in inject fluid channel, multistage fluid channel and output fluid channel structural representation；
Fig. 3 is the main section of the microinjection chip of the present invention；
The arrangement schematic diagram of each electrode in each microelectrode array in Tu4Shi lower floor quartz substrate；
Fig. 5 be the present invention upper strata quartz substrate in electrode structure schematic diagram；
Fig. 6 is the cell of present invention operating diagram in injection fluid channel.
In figure:
Lower floor's quartz substrate 1, miniflow channel layer 2, upper strata quartz substrate 3, PDMS block 4, injection needle 5, cell 6；
201, two grades of fluid channel 202 of one-level fluid channel, injection fluid channel 203, output fluid channel 204；
First microelectrode array the 11, second microelectrode array the 12, the 3rd microelectrode array the 13, the 4th microelectrode array 14,
5th microelectrode array 15；
Each parallel pole 1101,1102,1103,1104,1105 and 1106 in first microelectrode array；
Two fork-shaped electrode 1201,1202 in second microelectrode array；
Each parallel pole 1301,1302,1303,1304 in 3rd microelectrode array；
Each electrode 1401,1402,1403,1404 in 4th microelectrode array；
Each electrode 1501 in 5th microelectrode array, 1502,1503,1504,1505,1506,1507,1508,1509,
1510。
Detailed description of the invention
In conjunction with the accompanying drawings, the present invention is further detailed explanation.These accompanying drawings are the schematic diagram of simplification, only with
The basic structure of the illustration explanation present invention, therefore it only shows the composition relevant with the present invention.
Owing to current microinjection device did not the most adjust its position before cell is carried out microinjection, study table
Bright in intracytoplasmic sperm injection, oocyte first polar body is positioned at diverse location, sends out cell survival rate, Fertilized quality and embryo
Educate and all have a significant impact.Generally requirement of experiment first polar body is away from the operating position of entry needle, to reduce cells such as spindles
The injury of device.And this cell microinjection chip, cell microinjection device and method of work are carrying out micro-note to cell
Can adjust its attitude before penetrating, this carries out the survival rate of cell after micrurgy by being greatly improved.
Term is explained:
Dielectrophoresis: also referred to as two-dimensional electrophoresis, is neutral particle stress phenomenon of moving in non-uniform electric field.Comprise, often
Rule dielectrophoresis, row ripple dielectrophoresis and rotation dielectrophoresis.
Polydimethylsiloxane: (Polydimethylsioxane) is called for short PDMS, is a kind of the macromolecule organic silicon chemical combination
Thing.There is optical clear, and in the ordinary course of things it is considered to be inertia, nontoxic, nonflammable, there is good bio-compatibility.
Embodiment 1
As shown in figures 1 and 3, cell microinjection chip includes four parts, stack gradually the most from bottom to top containing phase
Answer lower floor's quartz substrate 1 of microelectrode array, miniflow channel layer 2, the upper strata quartz substrate 3 containing microelectrode array, microinjection
The PDMS block 4 of pin 5 and fixing injection needle；Injection needle 5 and PDMS block 4 is by microinjection needle handle and PDMS block
Aperture interference fit realize seal.
Concrete, cell microinjection chip includes: injection fluid channel 203 and multistage fluid channel, wherein said multistage micro-
Runner is suitable to screen cell, so that only one cell 6 enters in injection fluid channel 203, cell 6 is clashed into microinjection
The needle point of pin 5 is to carry out microinjection again after puncturing cell；And the center that the needle point of injection needle 5 is by upper strata quartz substrate
Hole is stretched in injection fluid channel 203.
As in figure 2 it is shown, described multistage fluid channel includes: at least two-stage fluid channel；I.e. one-level fluid channel 201 and two grades of miniflows
Road 202, and the width of flow path of next stage fluid channel is less than the width of flow path of upper level fluid channel；And described one-level fluid channel
Bottom is provided with the first microelectrode array 11, and this first microelectrode array 11 is suitable to drive after powered up cell suspension and micro-to two grades
Runner moves；The bottom of described two grades of fluid channel is provided with the second microelectrode array 12；When cell enters two grades from one-level fluid channel
After fluid channel, adjust cell by the second microelectrode array and enter hoverheight and the translational speed of injection fluid channel.
The width of flow path of two grades of fluid channel is more than the width of flow path of injection fluid channel, the width of flow path of described injection fluid channel
More than cell dia to be injected, and less than 2 times of cell dia to be injected.
The bottom of described injection fluid channel is provided with the 3rd microelectrode array 13, and top is provided with the 4th microelectrode array 14, with
And the 4th the center of microelectrode array be provided with described injection needle 5；Described 3rd microelectrode array be suitable to control cell to
The underface of injection needle is moved, and by described 4th microelectrode array capture cell so that it is it is positioned injection needle
Underface；And by, after the three, the 4th microelectrode array synergic adjustment cell attitudes, making cell move up or down；
When moving on cell, described microinjection is needled into cell, injects；When cell moves down, inject cell detachment micro-
Entry needle, and entered output fluid channel 204 by the 3rd microelectrode array by injecting cell.
The bottom of described output fluid channel 204 is provided with the 5th microelectrode array 15；And be energized at the 5th microelectrode array 15
After, make to inject cell output.
Described upper level fluid channel is horn mouth transition with the junction of lower two grades of fluid channel.
Described injection fluid channel, multistage fluid channel and output fluid channel are respectively positioned in miniflow channel layer, described first, second,
3rd and the 5th microelectrode array is distributed in lower floor's quartz substrate, and the 4th microelectrode array is distributed in upper strata quartz substrate.
As shown in Figure 4 and Figure 5, described first, the 3rd and the 5th microelectrode array is arranged at equal intervals by parallel strip electrodes
Form, and parallel strip electrodes is orthogonal with the moving direction of cell；Described second microelectrode array use two fork-shaped electrode fingers and
Become, and finger electrode is in being spacedly distributed；And in described 4th microelectrode array in each electrode above quartz substrate center
Centered by central hole, circumferential array obtains, 90 ° each other of the most each electrode, and extends to the central axis of injection needle.
The size of fluid channel and electrode can change in certain proportion according to the size of cell to be injected.
The relevant enforcement step of this cell microinjection chip will be discussed in detail in embodiment 3.
Embodiment 2
On the basis of embodiment 1, the present embodiment 2 additionally provides a kind of cell microinjection device.
Described cell microinjection device includes: for connecting the control of each microelectrode array in cell microinjection chip
Module；Described control module produce each microelectrode array corresponding control signal, with control cell from multistage fluid channel move to
Injection fluid channel is to inject cell location, pose adjustment, and will to inject cell from entry needle after injection completes
Export after disengaging.
Cell microinjection chip involved in the present embodiment 2 is as described in Example 1.
Embodiment 3
On the basis of embodiment 1 and embodiment 2, the present embodiment 3 additionally provides the work of a kind of cell microinjection device
Method.
The method of work of described cell microinjection device, comprises the steps:
Step S1, screens cell, makes only one cell enter in injection fluid channel；
Step S2, injects entering the cell injecting in fluid channel；And
Step S3, will inject cell output.
Described cell microinjection device is suitable for use with cell microinjection device as described in Example 2, and this is thin
The cell microinjection as described in Example 1 that cell microinjection chip used in born of the same parents' microinjection device is suitable for use with
Chip.
Cell is screened by described step S1, makes only one cell enter the concrete grammar bag in injection fluid channel
Include: with pipettor, suspension cell solution to be injected is injected in chip from one-level fluid channel；In cell microinjection device
Control module is suitable for use with to produce the multichannel sine wave signal generator of frequency and phase-adjustable to the first microelectrode array
In each parallel pole (1101,1102,1103,1104,1105 and 1106) apply after phase place differs the sinusoidal signal of 90 degree successively,
Produce inhomogeneous field in making one-level fluid channel 201, so make its inner cell can simultaneously by Conventional dielectric swimming power and
(the Conventional dielectric swimming field of force makes cell suspension in fluid channel to row ripple dielectrophoretic force, is made cellular water by the row ripple dielectrophoresis field of force
Put down to left movement)；When moving to horn mouth transition region under the effect of cell row ripple dielectrophoretic force in one-level fluid channel 201
In, the phase place stopping the sinusoidal signal on the first microelectrode array changes, and reduces cell and enters the mobile speed of two grades of fluid channel 202
Degree, plays the effect of buffering；The second microelectrode array (two fork-shaped electrodes 1201,1202) bottom two grades of fluid channel 202 again
Upper applying sinusoidal signal, makes the effect of cell dielectrophoretic force in two grades of fluid channel lower the hoverheight of ganglion cell and to it
Slow down, i.e. cell buffers under the effect of motional inertia in this region, and reduces mobile to the speed injecting fluid channel 203.
Described step S2 includes entering the method that the cell injecting in fluid channel injects:
The row ripple dielectrophoresis that parallel pole (1301,1302,1303 and 1304) induction in 3rd microelectrode array produces
Power can make cell 6 rotation forward or backward, continues to move horizontally simultaneously；The sense of rotation of cell 6 sees Fig. 6.The most corresponding with it
Top the 4th microelectrode array in each electrode (1401,1402,1403 and 1404) Conventional dielectric swim under produce dielectric
Swimming power can be controlled cell 6 be in all the time by attraction and capture cell 6 (its force trapping F sees the citing in Fig. 6 and illustrates)
The underface of microinjection pin；Row ripple dielectrophoretic force and the Conventional dielectric of the 4th microelectrode array that 3rd microelectrode array produces are swum
The coupling of both power can be with manipulation cell attitude and kinestate, it may be assumed that hover over the underface of microinjection pin 5, can make
Cell spins and vertically moves up and down；Described cell microinjection device is suitable to according to cell movement tendency now, and with in advance
The cell movement direction and goal position versus of phase, carries out corresponding reaction type regulation, i.e. by control to signal frequency and voltage
The signal frequency of signal generator processed generation and voltage control frequency and the phase place of its dielectrophoretic force produced indirectly, so that carefully
Born of the same parents' spatial attitude in this region is adjusted；Concrete, each electrode in the 4th microelectrode array on upper strata quartz substrate 3
(1401,1402,1403 and 1404) produce non-uniform electric field, such as but not limited to apply the phase place of signal be 0 °, 180 °,
0 °, 180 ° time, the Conventional dielectric swimming power of induction can control cell and straight up or moves downward；Have continuously applying signal
During phase place (such as 0 °, 90 °, 180 °, 270 °) of equal difference, the rotation dielectrophoretic force that induction produces can control cell in horizontal plane
Rotate.After cell pose adjustment is good, regulate pattern and the frequency of sinusoidal signal on the three, the 4th microelectrode arrays so that it is lure
Artificial deliviery raw Conventional dielectric swimming power makes cell move straight up, after cell is thrust by microinjection pin 5, by being connected with microinjection pin
Accurate micro syringe pump by allogenic material solution to be injected inject cell interior, after cell completes microinjection, regulate the three, the 4th
The frequency of sinusoidal signal on microelectrode array, the Conventional dielectric swimming power that its induction produces make to inject cell move straight down and
Depart from microinjection pin, then regulate four parallel poles (1301,1302,1303 and 1304) in the 3rd microelectrode array and produce non-even
Highfield, the row ripple dielectrophoretic force that induction produces can control the cell of microinjection and move in the horizontal direction to output miniflow
Road 204.
The method injecting cell output is included by described step S3:
When cell enters after fluid channel 204, each electrode in regulation the 5th microelectrode array (1501,1502,1503,1504,
1505,1506,1507,1508,1509 and 1510) phase and frequency of the AC signal on (i.e. applies phase 90 degree
Sinusoidal signal, and regulate the frequency of sinusoidal signal), made the cell of microinjection be expert at ripple dielectrophoretic force effect under move to
The output port of output fluid channel 204.And so forth, the microinjection operation of remaining cell in fluid channel can be completed.
Optionally, in each microelectrode array the quantity of respective electrode such as but not limited to listed in embodiment 1 to embodiment 3
The quantity lifted, such as 8 or 12 or 16.
With the above-mentioned desirable embodiment according to the present invention for enlightenment, by above-mentioned description, relevant staff is complete
Entirely can carry out various change and amendment in the range of without departing from this invention technological thought.The technology of this invention
The content that property scope is not limited in description, it is necessary to determine its technical scope according to right.
Claims (10)
1. a cell microinjection chip, it is characterised in that including:
Injection fluid channel and multistage fluid channel, wherein
Described multistage fluid channel is suitable to screen cell, so that only one cell enters in injection fluid channel, carries out micro-
Injection.
Cell microinjection chip the most according to claim 1, it is characterised in that
Described multistage fluid channel includes: at least two-stage fluid channel；I.e.
One-level fluid channel and two grades of fluid channel, and the width of flow path of next stage fluid channel is less than the runner width of upper level fluid channel
Degree；And
The bottom of described one-level fluid channel is provided with the first microelectrode array, and this first microelectrode array is suitable to drive after powered up carefully
Born of the same parents suspend and move to two grades of fluid channel；
The bottom of described two grades of fluid channel is provided with the second microelectrode array；
When cell is after one-level fluid channel enters two grades of fluid channel, adjust cell by the second microelectrode array and enter injection miniflow
The hoverheight in road and translational speed.
Cell microinjection chip the most according to claim 2, it is characterised in that
The bottom of described injection fluid channel is provided with the 3rd microelectrode array, and top is provided with the 4th microelectrode array, and the 4th is micro-
The center of electrod-array is provided with injection needle；
Described 3rd microelectrode array is suitable to control cell and moves to the underface of injection needle, and by described 4th micro-electricity
Pole array capture cell so that it is be positioned the underface of injection needle；And
After the three, the 4th microelectrode array synergic adjustment cell attitudes, cell is made to move up or down；
When moving on cell, described microinjection is needled into cell, injects；
When cell moves down, inject cell detachment injection needle, and entered injecting cell by the 3rd microelectrode array
Output fluid channel.
Cell microinjection chip the most according to claim 3, it is characterised in that the bottom of described output fluid channel is provided with
5th microelectrode array；And
After the 5th microelectrode array energising, make to inject cell output.
Cell microinjection chip the most according to claim 4, it is characterised in that described upper level fluid channel and lower two grades
The junction of fluid channel is horn mouth transition.
Cell microinjection chip the most according to claim 5, it is characterised in that
Described injection fluid channel, multistage fluid channel and output fluid channel are positioned in miniflow channel layer, and
The upper and lower quartz substrate that has been covered each by of described miniflow channel layer, described first, second, third and the 5th microelectrode array
Being distributed in lower floor's quartz substrate, the 4th microelectrode array is distributed in upper strata quartz substrate.
Cell microinjection chip the most according to claim 6, it is characterised in that
Described first, the 3rd and the 5th microelectrode array is arranged at equal intervals by parallel pole and forms, and
Parallel pole is orthogonal with the moving direction of cell；
Described second microelectrode array uses two fork-shaped electrode fingers to form；And
In described 4th microelectrode array, each electrode is at the spaced 90 ° of angles of circumferencial direction, and to the center of injection needle
Extend at axle.
8. a cell microinjection device, it is characterised in that including:
For connecting the control module of each microelectrode array in cell microinjection chip；
Described control module produces the corresponding control signal of each microelectrode array, moves to note from multistage fluid channel controlling cell
Penetrate fluid channel so that cell location, pose adjustment are injected, and to be taken off injecting cell from entry needle after injection completes
From rear output.
9. a method of work for cell microinjection device, comprises the steps:
Step S1, screens cell, makes only one cell enter in injection fluid channel；
Step S2, injects entering the cell injecting in fluid channel；And
Step S3, will inject cell output.
Method of work the most according to claim 9, it is characterised in that
Cell microinjection chip used in described cell microinjection device is suitable for use with such as any one of claim 4-7
Described cell microinjection chip.
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| CN201610615682.6A CN106244416B (en) | 2016-07-29 | 2016-07-29 | Cell microinjection chip, injection device and its working method based on dielectrophoresis |
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| CN201610615682.6A CN106244416B (en) | 2016-07-29 | 2016-07-29 | Cell microinjection chip, injection device and its working method based on dielectrophoresis |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108359577A (en) * | 2018-01-29 | 2018-08-03 | 燕山大学 | Zebra fish-egg rotation micro OS based on micro fluid dynamcis and control method |
| CN109706054A (en) * | 2019-01-04 | 2019-05-03 | 哈尔滨工业大学 | The automatic conveying of zebra fish juvenile fish and the micro runner device of adjustment posture |
| CN111909839A (en) * | 2019-05-08 | 2020-11-10 | 香港城市大学深圳研究院 | System for delivering substances into cells, master control device, microcontroller and method |
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| CN111909839A (en) * | 2019-05-08 | 2020-11-10 | 香港城市大学深圳研究院 | System for delivering substances into cells, master control device, microcontroller and method |
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