CN103396944A - Electroporation chip for cell transfection and making method thereof - Google Patents
Electroporation chip for cell transfection and making method thereof Download PDFInfo
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- CN103396944A CN103396944A CN2013103086442A CN201310308644A CN103396944A CN 103396944 A CN103396944 A CN 103396944A CN 2013103086442 A CN2013103086442 A CN 2013103086442A CN 201310308644 A CN201310308644 A CN 201310308644A CN 103396944 A CN103396944 A CN 103396944A
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Abstract
The invention discloses an electroporation chip for cell transfection and a making method thereof. The electroporation chip comprises a base plate and a three-dimensional electrode arranged on the surface of the base plate, wherein the surface deviating from the base plate, of the three-dimensional electrode is a top face, the surface facing to the base plate, of the three-dimensional electrode is a bottom surface, and the surface located between the top face and the bottom surface is a side face; and the side face is connected with the top face through a cambered surface. Since the three-dimensional electrode with large thickness is adopted on the electroporation chip, compared with the traditional two-dimensional electrodes, the thickness in the direction vertical to the base plate is large, so that the distribution region of an electric field is larger and cell transfection efficiency is beneficial to improvement; and moreover, the bottom surface of the three-dimensional electrode is connected with the side face through the cambered surface to avoid the tip effect due to edges and corners of the electrode, ensure good uniformity of the electric field, guarantee the survival rate of cells and further improve the cell transfection efficiency.
Description
Technical field
The present invention relates to the cell transfecting technical field, more particularly, relate to a kind of chip of electroporation for cell transfecting and preparation method thereof.
Background technology
Cell transfecting refers to the process of allogenic material transfered cell, is the prerequisite of gene therapy and RESEARCH ON CELL-BIOLOGY.At present, can realize cell transfecting by modes such as chemical transfection, virus transfection, particle gun, microinjection and electroporations.Wherein, electroporation, with low cytotoxicity, low cost and transfection efficiency advantages of higher, becomes the topmost use-pattern of present cell transfecting.
Electroporation is to make cell surface produce recoverable micropore by pulsed electrical field, thereby by foreign aid's material transfered cell.When carrying out electroporation, be generally to adopt the electroporation chip that is provided with electrode to provide pulsed electrical field for the cell cultures vessel.
Size and the inhomogeneity quality of electroporation chip electric field region are the key factors that affects cell transfecting efficiency.How, when improving electroporation chip electric field region, guarantee that the homogeneity of electric field is a current problem urgently to be resolved hurrily.
Summary of the invention
For solving the problems of the technologies described above, the invention provides a kind of chip of electroporation for cell transfecting and preparation method thereof, the electric field region of described electroporation is larger, and the homogeneity of electric field is better.
For achieving the above object, the invention provides following technical scheme:
A kind of chip of electroporation for cell transfecting, described electroporation chip comprises:
Substrate;
Be arranged on the three-diemsnional electrode of described substrate surface;
Wherein, the surface that described three-diemsnional electrode deviates from described substrate is end face, towards the surface of described substrate, is bottom surface, and the surface between described end face and described bottom surface is side; Between described side and described end face, for cambered surface, be connected.
Preferably, in above-mentioned electroporation chip, the thickness range of described three-diemsnional electrode is 1 μ m-1000 μ m, comprises endpoint value;
Described thickness is the distance of described bottom surface to described end face.
Preferably, in above-mentioned electroporation chip, the width range of three-diemsnional electrode is 10 μ m-1000 μ m, comprises endpoint value;
Described width is that described three-diemsnional electrode is in the live width perpendicular on described three-diemsnional electrode bearing of trend.
Preferably, in above-mentioned electroporation chip, described three-diemsnional electrode is golden three-diemsnional electrode or copper three-diemsnional electrode or tin three-diemsnional electrode or alloy three-dimensional electrode or conductive polymers three-diemsnional electrode.
Preferably, in above-mentioned electroporation chip, described three-diemsnional electrode is straight line.
Preferably, in above-mentioned electroporation chip, described three-diemsnional electrode is annulus.
Preferably, in above-mentioned electroporation chip, the end of described three-diemsnional electrode is cambered surface.
The present invention also provides a kind of making method of electroporation chip, and the method comprises:
Formation cross-section is the three-diemsnional electrode of rectangle on substrate, wherein, the surface that described three-diemsnional electrode deviates from described substrate is end face, towards the surface of described substrate, it is bottom surface, surface between described end face and described bottom surface is side, and described cross section is the plane perpendicular to described three-diemsnional electrode bearing of trend;
Described three-diemsnional electrode is carried out to smooth treatment, between the end face after making to process and side, for cambered surface, connect.
Preferably, in above-mentioned making method, described on substrate Formation cross-section be that the three-diemsnional electrode of rectangle is:
By printed circuit board technology Formation cross-section on described substrate, it is the three-diemsnional electrode of rectangle.
Preferably, in above-mentioned making method, described on substrate Formation cross-section be that the three-diemsnional electrode of rectangle is:
At described substrate surface, form mask layer, described mask layer is provided with rectangular recess;
Deposits conductive material in described rectangular recess, Formation cross-section are the three-diemsnional electrode of rectangle;
Remove described mask layer.
Preferably, in above-mentioned making method, described smooth treatment is:
In described cross section, be evaporation conduction transition layer on the three-diemsnional electrode surface of rectangle, between the end face that makes to cover the three-diemsnional electrode after described conduction transition layer and side, realize that cambered surface is connected.
Preferably, in above-mentioned making method, described smooth treatment is:
The corner angle that are the three-diemsnional electrode of rectangle to described cross section corrode, thereby make between its end face and side, realize that cambered surface connects.
Preferably, in above-mentioned making method, described smooth treatment is:
The three-diemsnional electrode that is rectangle to described cross section carries out high temperature melting, by the surface tension of the three-diemsnional electrode after fusing, eliminates the corner angle between former end face and side;
Three-diemsnional electrode after fusing is cured to typing, forms the three-diemsnional electrode that between side and end face, cambered surface is connected.
The present invention also provides the making method of another kind of electroporation chip, and the method comprises:
On substrate, fix the mould of a three-diemsnional electrode, the surface that described mould is relative with described substrate is provided with groove, between the bottom surface of described groove and side, for cambered surface, be connected, wherein, described bottom surface is the surface that described groove deviates from described substrate, and described side is the surface between described bottom surface and described substrate;
In described groove, inject liquid electro-conductive material;
Make the electro-conductive material solidifying and setting in described groove, remove described mould, form three-diemsnional electrode.
From technique scheme, can find out, electroporation for cell transfecting chip provided by the present invention adopts the larger three-diemsnional electrode of thickness, it is with respect to traditional two-dimensional electrode, thickness on the direction perpendicular to substrate is larger, the distributed areas of electric field are larger, are conducive to improve the transfection efficiency of cell; And between the bottom surface of described three-diemsnional electrode and side, for cambered surface is connected, avoided the point effect that exists corner angle to cause due to electrode, made the homogeneity of electric field better, guaranteed the survival rate of cell, further improved the transfection efficiency of cell.The making method of electroporation chip provided by the invention can be made above-mentioned electroporation chip, and technique is simple.
The accompanying drawing explanation
In order to be illustrated more clearly in the embodiment of the present invention or technical scheme of the prior art, below will the accompanying drawing of required use in embodiment or description of the Prior Art be briefly described, apparently, accompanying drawing in the following describes is only some embodiments of the present invention, for those of ordinary skills, under the prerequisite of not paying creative work, can also obtain according to these accompanying drawings other accompanying drawing.
The structural representation of a kind of chip of electroporation for cell transfecting that Fig. 1 provides for the embodiment of the present invention;
Fig. 2 is the simulation result schematic diagram of the electric field distribution of two-dimensional electrode;
Fig. 3 is the simulation result schematic diagram of electric field distribution of the three-diemsnional electrode of square-section;
Fig. 4 is the simulation result schematic diagram of the electric field distribution of the three-diemsnional electrode of employing cambered surface connection;
Fig. 5 is three kinds of electrodes distribution map of the electric field at 1 micron place above it;
The electrode structure schematic diagram of a kind of electroporation chip that Fig. 6 provides for the embodiment of the present invention;
The electrode structure schematic diagram of the another kind of electroporation chip that Fig. 7 provides for the embodiment of the present invention;
The principle schematic of a kind of cell transfecting that Fig. 8 provides for the embodiment of the present invention;
The principle schematic of the another kind of cell transfecting that Fig. 9 provides for the embodiment of the present invention;
The schematic flow sheet of the making method of a kind of electroporation chip that Figure 10 provides for the embodiment of the present invention;
A kind of schematic flow sheet that forms the three-diemsnional electrode of square-section that Figure 11-Figure 13 provides for the embodiment of the present invention;
The schematic flow sheet of the making method of the another kind of electroporation chip that Figure 14 embodiment of the present invention provides;
The structural representation of a kind of mould for the preparation of the electroporation chip that Figure 15 provides for the embodiment of the present invention.
Embodiment
Just as described in the background section, how, when improving electroporation chip electric field region, guarantee that the homogeneity of electric field is a current problem urgently to be resolved hurrily.
The contriver studies discovery, can adopt the three-diemsnional electrode with larger thickness to increase electric field region, simultaneously by electrode structure is set, makes between electrode phase cross surface and connect by cambered surface, avoid the point effect that exists corner angle to cause due to electrode, guarantee the homogeneity that electric field divides.
Based on above-mentioned research, the invention provides a kind of chip of electroporation for cell transfecting, this electroporation chip comprises:
Substrate;
Be arranged on the three-diemsnional electrode of described substrate surface;
Wherein, the surface that described three-diemsnional electrode deviates from described substrate is end face, towards the surface of described substrate, is bottom surface, and the surface between described end face and described bottom surface is side; Between described side and described end face, for cambered surface, be connected.
Electroporation for cell transfecting chip provided by the present invention adopts the larger three-diemsnional electrode of thickness, it is with respect to traditional two-dimensional electrode, thickness on the direction perpendicular to substrate is larger, and the distributed areas of electric field are larger, is conducive to improve the transfection efficiency of cell; And between the bottom surface of described three-diemsnional electrode and side, for cambered surface is connected, avoided the point effect that exists corner angle to cause due to electrode, made the homogeneity of electric field better, guaranteed the survival rate of cell, further improved the transfection efficiency of cell.
It is more than the application's core concept, below in conjunction with the accompanying drawing in the embodiment of the present invention, the technical scheme in the embodiment of the present invention is clearly and completely described, obviously, described embodiment is only the present invention's part embodiment, rather than whole embodiment.Based on the embodiment in the present invention, those of ordinary skills, not making under the creative work prerequisite the every other embodiment that obtains, belong to the scope of protection of the invention.
A lot of details have been set forth in the following description so that fully understand the present invention, but the present invention can also adopt other to be different from alternate manner described here and implement, those skilled in the art can be in the situation that do similar popularization without prejudice to intension of the present invention, so the present invention is not subjected to the restriction of following public specific embodiment.
Secondly, the present invention is described in detail in conjunction with schematic diagram, when the embodiment of the present invention is described in detail in detail; for ease of explanation; the schematic diagram of indication device structure can be disobeyed general ratio and be done local the amplification, and described schematic diagram is example, and it should not limit the scope of protection of the invention at this.The three-dimensional space that should comprise in addition, length, width and height in actual fabrication.
Embodiment mono-
Based on above-mentioned thought, the present embodiment provides a kind of chip of electroporation for cell transfecting, with reference to figure 1, comprising: substrate 1 and be positioned at the lip-deep three-diemsnional electrode 2 of described substrate 1.
Fig. 1 is the sectional view along described three-diemsnional electrode bearing of trend, and wherein, described three-diemsnional electrode 2 is bottom surface with the surface that described substrate contacts fully, and the surface relative with described bottom surface is end face, and the surface between described bottom surface and end face is side.Between the end face of the described three-diemsnional electrode of the present embodiment and side, for cambered surface, be connected.
In the present embodiment, the thickness range of described three-diemsnional electrode 2 is 1 μ m-1000 μ m, comprises endpoint value, and described thickness is the distance of described bottom surface to described end face, the i.e. length value on vertical direction in Fig. 2.The width range of described three-diemsnional electrode 2 is 10 μ m-1000 μ m, comprises endpoint value, described width be described three-diemsnional electrode in the live width perpendicular on described three-diemsnional electrode bearing of trend, i.e. the length value of horizontal direction in Fig. 1.Described three-diemsnional electrode can adopt the preparations such as metal (as gold, copper, tin etc.), metal alloy or conductive polymers.
The thickness of traditional electroporation chip electrode is not more than the two-dimensional electrode of 0.5 μ m.Due to its thinner thickness, make electric field region less, and, because point effect is stronger, cause field uniformity poor.
And only increase the thickness of electrode, such as the thickness that electrode is set more than 1 μ m, cross section is rectangle, although can increase electric field in the distributed areas perpendicular on orientation substrate, still can there be stronger point effect in the electrode corner angle.
The present embodiment, when increasing thickness of electrode, makes between electrode end face and side and is connected by cambered surface, has greatly weakened point effect, has guaranteed the homogeneity of electric field.
Below by concrete analysis of simulation experiment two-dimensional electrode, cross section, be the three-diemsnional electrode of rectangle and adopt the electric field region of the three-diemsnional electrode of cambered surface connection to distribute.
While adopting the electric field distribution of limit meta software Comsol emulation Different electrodes, the material of three kinds of electrodes of design is all identical, and width all is set to 150 μ m, and spacing all is set to 250 μ m.For two-dimensional electrode, it is 0.5 μ m that its thickness is set, and cross section is that the three-diemsnional electrode of rectangle and the thickness of the three-diemsnional electrode that the employing cambered surface connects all are set to 40 μ m, and the voltage that applies of all electrodes is 20V.Simulation result as Figure 2-Figure 5.
In the 0.2mm zone, the two-dimensional electrode electric field is greater than 5 * 10 above substrate
4The area percentage that the zone of V/m accounts for overall area (substrate top 0.2mm take interior zone) as 32%(as shown in Figure 2).Cross section is that the three-diemsnional electrode of rectangle and the electric field that adopts the three-diemsnional electrode that cambered surface connects are greater than 5 * 10
4The area percentage that the zone of V/m accounts for overall area all greater than 60%(referring to Fig. 3 and Fig. 4).As can be known by Fig. 2-Fig. 4, with respect to two-dimensional electrode, the electric field region of three-diemsnional electrode obviously increases.
At each electrode top face 1 μ m place of Fig. 2-Fig. 4, do respectively sea line, gather separately strength of electric field value on sea line, draw the one dimension distribution map of the electric field, as shown in Figure 5.
With reference to figure 5, the electric field distribution at 1 μ m place is as can be known above it by three kinds of electrodes, two-dimensional electrode point effect maximum, two peak values of sampling are respectively 5.31V/m, 5.39V/m, secondly be the three-diemsnional electrode of square-section, two peak electric field of its sampling are 2.81V/m, 2.78V/m, the three-diemsnional electrode for adopting cambered surface to connect that point effect is the most weak, two peak electric field of its sampling are 1.38V/m, 1.37V/m, and most of strength of electric field of three kinds of electrodes are near 0.46V/m.As shown in Figure 5, the homogeneity of the three-diemsnional electrode electric field of employing cambered surface connection is better.
The three-diemsnional electrode of the described electroporation chip of the present embodiment can be straight line, with reference to figure 6, when described three-diemsnional electrode 2 is straight line, the electrode structure of described electroporation chip is the finger-inserting type electrode, the positive pole "+" that is the electroporation chip consists of the three-diemsnional electrode 2 of a plurality of linearities, form anodal array, negative pole "-", by the three-diemsnional electrode 2 of a plurality of linearities, forms cathode array.The three-diemsnional electrode 2 of anodal "+" linearity is spaced apart with the three-diemsnional electrode 2 of negative pole "-" linearity.
General, the cell cultures chamber is cylindrical, so described positive pole "+" and negative pole "-" all adopt circular arc connection section 3 three-diemsnional electrode 2 connections of linearity separately.At the electroporation chip for electrode structure shown in Figure 6, in order further to reduce point effect, the free end of the three-diemsnional electrode of linearity is cambered surface, and namely to deviate from an end of described circular arc connection section 3 be cambered surface to the three-diemsnional electrode 2 of linearity.
The three-diemsnional electrode of the described electroporation chip of the present embodiment can be annulus, with reference to figure 7, when described three-diemsnional electrode 2 is annulus, the electrode structure of described electroporation chip is the nested electrode, the positive pole "+" that is the electroporation chip consists of a plurality of circular three-diemsnional electrodes 2, form anodal array, negative pole "-" consists of a plurality of circular three-diemsnional electrodes 2, forms cathode array.At this moment, circular mutually nested being alternately distributed of three-diemsnional electrode 2 of anodal "+" circular three-diemsnional electrode 2 and negative pole "-".
In the electroporation chip of electrode structure shown in Figure 7, mutually nested with cathode array in order to realize anodal array, the circular three-diemsnional electrode 2 that is positioned at innermost layer can also can be for being provided with the part annulus of opening for complete circle, and other annulus are the part annulus that is provided with opening.Equally, in order further to reduce point effect, for the part annulus of opening, two free ends of opening part are cambered surface.For circular three-diemsnional electrode, the cross section of its bearing of trend is the cross section of tangent line.
Identical with existing electroporation chip, while adopting the described electroporation chip of the present embodiment to carry out cell transfecting, must provide the space of a solution carrying, hold the electroporation system, described system comprises: cell, band transfection material and solution etc.General, adopt the orifice plate that is provided with a plurality of cavernous structures to carry out cell transfecting.
As shown in Figure 8, the described electroporation chip of the present embodiment, when carrying out cell transfecting, can be arranged on the below of electroporation system.In the bottom of the through-hole structure of orifice plate 4 by the described electroporation chip of glue bond the present embodiment.Described orifice plate 4 can adopt by polydimethylsiloxane (PDMS) or polymethylmethacrylate (PMMA) preparation.
In the transfection system of mode shown in Figure 8, for attached cell, can first be added to interior the cultivation 24 hours of through hole of the orifice plate 4 that is provided with the described electroporation chip of the present embodiment, after it is adherent, add again the electroporation damping fluid and treat the transfection material.Apply the electroporation pulse then for the electrod-array of each hole or duct bottom, cell is carried out to electroporation, thereby will treat that the transfection material imports cultivation in the cell of bottom, hole.With cell culture medium displacement electroporation damping fluid, electroporation chip be placed in cell culture incubator continue cultivate, 24 to 48 hour after the transfection effect of observation of cell thereafter.Electroporation for the cell of suspended state is similar with it, and attached cell or the bacterium of suspension cell or digestion rear-suspending can directly mix and be added in described through hole with the electroporation system.After applying electricimpulse, can be directly toward dripping a large amount of cell culture mediums in chip, by cell in-situ, cultivate, or by the cell after electroporation outstanding be that sucking-off is added in the Tissue Culture Plate that is added with in advance a large amount of substratum and cultivates.Whether the electroporation damping fluid in cell suspension needs to remove and be determined on a case-by-case basis by centrifuging.
As shown in Figure 9, the described electroporation chip of the present embodiment, when carrying out cell transfecting, can be arranged on the top of electroporation system.
Can increase baffle plate 5 in the both sides of substrate 1, then electrode be immersed and contains cell and treat in the orifice plate of transfection material, described baffle plate 5 guarantees that the electroporation chip can directly not contact the bottom of the through hole of orifice plate 4, guarantees that cell is not extruded.Thereafter in electroporation process Fig. 8, process is similar.The method can be applied to cell under adherent state and suspended state and transfection or the conversion of bacterium.Maximum advantage is the orifice-plate type cell cultures compatibility of perforation procedure and routine.
The described electroporation chip of the present embodiment can be used for transfered cells such as exogenous nucleic acid (DNA, RNA, siRNA etc.), protein, small-molecule drugs.
As can be known by foregoing description, have the electroporation chip that adopts the three-diemsnional electrode that cambered surface connects, when carrying out cell transfecting, the electric field distribution zone is large, and the good uniformity of electric field, can effectively improve cell transfecting efficiency.
Embodiment bis-
The present embodiment provides a kind of making method of electroporation chip, and the electrode of described electroporation chip is the three-diemsnional electrode that comprises that the described employing cambered surface of above-described embodiment connects, and with reference to Figure 10, this making method comprises:
Step S1: Formation cross-section is the three-diemsnional electrode of rectangle on substrate.
The surface that described three-diemsnional electrode deviates from described substrate is end face, towards the surface of described substrate, is bottom surface, and the surface between described end face and described bottom surface is side, and described cross section is the cross section perpendicular to described three-diemsnional electrode bearing of trend.
Can be the three-diemsnional electrode of rectangle by printed circuit board technology (PCB technique) Formation cross-section on described substrate.The PCB process quilt is widely used in the processing of the electronic circuit field of suitability for industrialized production, and cost is low, but rapid shaping processing also can be mass.The precision of the thickness of electrode that PCB technique is made has reached tens microns, can be applied to make the required electrod-array of electroporation chip.
Can be also the three-diemsnional electrode of rectangle by electrochemical plating Formation cross-section on described substrate, comprise:
Step S11: with reference to Figure 11, on substrate 1 surface, form mask layer 6, described mask layer is provided with rectangular recess 7, namely on substrate 1, forms the mask layer with the groove that is complementary with electrode structure.The cross section of described rectangular recess 7 fingers on the groove bearing of trend is rectangle.
Step S12: with reference to Figure 12, in the interior deposits conductive material of above-mentioned groove 7, Formation cross-section is the three-diemsnional electrode 2' of rectangle.
Step S13: with reference to Figure 13, remove described mask layer.
Step S2: described three-diemsnional electrode is carried out to smooth treatment, realize between the end face after making to process and side that cambered surface connects.
Can, by in above-mentioned cross section, being evaporation conduction transition layer on the three-diemsnional electrode surface of rectangle, realize being the smooth treatment of the three-diemsnional electrode of rectangle to described cross section.Unshielded free evaporation is directly carried out on the three-diemsnional electrode surface that in described cross section is namely rectangle, on its surface, forms one deck transition layer, between the end face that makes the three-diemsnional electrode after vapor deposition treatment and side, realizes that cambered surface is connected.
Described transition layer be good biocompatibility electrode materials (as can adopt gold as described in transition layer), can avoid like this in step S1 adopting the injury of the virose electro-conductive material of cell tool (as metallic copper) to cell.
Also can realize to described cross section being the smooth treatment of the three-diemsnional electrode of rectangle by etching process.Adopting the described cross section of corrosive fluid corrosion is the corner angle of the three-diemsnional electrode of rectangle, makes between end face after corrosion and side and realizes that cambered surface connects.This mode need to form in step S1 described cross section be the three-diemsnional electrode of rectangle adopt to the cell hypotoxicity, with cell compatibility preferably electro-conductive material be prepared from.
Can also realize to described cross section being the smooth treatment of the three-diemsnional electrode of rectangle by remelting processing.At first, need to carry out high temperature melting to the three-diemsnional electrode that above-mentioned cross section is rectangle, by the surface tension of the three-diemsnional electrode after fusing, eliminate the corner angle between former end face and side.Then, the three-diemsnional electrode after fusing is cured to typing, forms the three-diemsnional electrode that between side and end face, cambered surface is connected.The method be applicable to adopt in step S1 low melting point, to the cell hypotoxicity, with cell compatibility preferably the cross section that is prepared from of electro-conductive material be the smooth treatment of the three-diemsnional electrode of rectangle.
As can be known by foregoing description, the described making method of the present embodiment, the Technology by routine have the electroporation chip of the three-diemsnional electrode that cambered surface connects in can Preparation Example one, and technique is simple.The electroporation chip electric field region of preparation is larger, and the homogeneity of electric field is better.
Embodiment tri-
The present embodiment provides the making method of another kind of electroporation chip, and the electrode of described electroporation chip is the three-diemsnional electrode that comprises that the described employing cambered surface of above-described embodiment connects, and with reference to Figure 14, this making method comprises:
Step S01: on substrate, fix the mould of a three-diemsnional electrode, the surface that described mould is relative with described substrate is provided with groove.
The shape that has the three-diemsnional electrode that cambered surface is connected in the shape of described groove and embodiment mono-is complementary, and between the bottom surface of described groove and side, for cambered surface, is connected.Wherein, described bottom surface is the surface that described groove deviates from described substrate, and described side is the surface between described bottom surface and described substrate.
With reference to Figure 15, described mould comprises: body 8; Be arranged on the groove 9 on 8 one surfaces of described body.Step S02: inject liquid electro-conductive material in described groove.
Step S03: make the electro-conductive material solidifying and setting in described groove, remove described mould, form three-diemsnional electrode.
The electroporation chip that adopts the described making method shape of the present embodiment to prepare, technique is simple, and the electroporation chip electric field region of preparation is larger, and the homogeneity of electric field is better.
To the above-mentioned explanation of the disclosed embodiments, make professional and technical personnel in the field can realize or use the present invention.Multiple modification to these embodiment will be apparent for those skilled in the art, and General Principle as defined herein can be in the situation that do not break away from the spirit or scope of the present invention, realization in other embodiments.Therefore, the present invention will can not be restricted to these embodiment shown in this article, but will meet the widest scope consistent with principle disclosed herein and features of novelty.
Claims (14)
1. the chip of the electroporation for cell transfecting, is characterized in that, comprising:
Substrate;
Be arranged on the three-diemsnional electrode of described substrate surface;
Wherein, the surface that described three-diemsnional electrode deviates from described substrate is end face, towards the surface of described substrate, is bottom surface, and the surface between described end face and described bottom surface is side; Between described side and described end face, for cambered surface, be connected.
2. electroporation chip according to claim 1, is characterized in that, the thickness range of described three-diemsnional electrode is 1 μ m-1000 μ m, comprises endpoint value;
Described thickness is the distance of described bottom surface to described end face.
3. electroporation chip according to claim 1, is characterized in that, the width range of three-diemsnional electrode is 10 μ m-1000 μ m, comprises endpoint value;
Described width is that described three-diemsnional electrode is in the live width perpendicular on described three-diemsnional electrode bearing of trend.
4. electroporation chip according to claim 1, is characterized in that, described three-diemsnional electrode is golden three-diemsnional electrode or copper three-diemsnional electrode or tin three-diemsnional electrode or alloy three-dimensional electrode or conductive polymers three-diemsnional electrode.
5. electroporation chip according to claim 1, is characterized in that, described three-diemsnional electrode is straight line.
6. electroporation chip according to claim 1, is characterized in that, described three-diemsnional electrode is annulus.
7. electroporation chip according to claim 1, is characterized in that, the end of described three-diemsnional electrode is cambered surface.
8. the making method of an electroporation chip, is characterized in that, comprising:
Formation cross-section is the three-diemsnional electrode of rectangle on substrate, wherein, the surface that described three-diemsnional electrode deviates from described substrate is end face, towards the surface of described substrate, it is bottom surface, surface between described end face and described bottom surface is side, and described cross section is the plane perpendicular to described three-diemsnional electrode bearing of trend;
Described three-diemsnional electrode is carried out to smooth treatment, between the end face after making to process and side, for cambered surface, connect.
9. making method according to claim 8, is characterized in that, described on substrate Formation cross-section be that the three-diemsnional electrode of rectangle is:
By printed circuit board technology Formation cross-section on described substrate, it is the three-diemsnional electrode of rectangle.
10. making method according to claim 8, is characterized in that, described on substrate Formation cross-section be that the three-diemsnional electrode of rectangle is:
At described substrate surface, form mask layer, described mask layer is provided with rectangular recess;
Deposits conductive material in described rectangular recess, Formation cross-section are the three-diemsnional electrode of rectangle;
Remove described mask layer.
11. the described making method of according to claim 8-10 any one, is characterized in that, described smooth treatment is:
In described cross section, be evaporation conduction transition layer on the three-diemsnional electrode surface of rectangle, between the end face that makes to cover the three-diemsnional electrode after described conduction transition layer and side, realize that cambered surface is connected.
12. the described making method of according to claim 8-10 any one, is characterized in that, described smooth treatment is:
The corner angle that are the three-diemsnional electrode of rectangle to described cross section corrode, thereby make between its end face and side, realize that cambered surface connects.
13. the described making method of according to claim 8-10 any one, is characterized in that, described smooth treatment is:
The three-diemsnional electrode that is rectangle to described cross section carries out high temperature melting, by the surface tension of the three-diemsnional electrode after fusing, eliminates the corner angle between former end face and side;
Three-diemsnional electrode after fusing is cured to typing, forms the three-diemsnional electrode that between side and end face, cambered surface is connected.
14. the making method of an electroporation chip, is characterized in that, comprising:
On substrate, fix the mould of a three-diemsnional electrode, the surface that described mould is relative with described substrate is provided with groove, between the bottom surface of described groove and side, for cambered surface, be connected, wherein, described bottom surface is the surface that described groove deviates from described substrate, and described side is the surface between described bottom surface and described substrate;
In described groove, inject liquid electro-conductive material;
Make the electro-conductive material solidifying and setting in described groove, remove described mould, form three-diemsnional electrode.
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