CN1699989A - Method for realizing low operating voltage electrophoresis and chips therefor - Google Patents
Method for realizing low operating voltage electrophoresis and chips therefor Download PDFInfo
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- CN1699989A CN1699989A CN 200510020641 CN200510020641A CN1699989A CN 1699989 A CN1699989 A CN 1699989A CN 200510020641 CN200510020641 CN 200510020641 CN 200510020641 A CN200510020641 A CN 200510020641A CN 1699989 A CN1699989 A CN 1699989A
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Abstract
The invention relates to a method and chips for working voltage electrophoresis, wherein it posits electrode array on micro-separator tube of electrophoresis chip, and posits auxiliary low-voltage on each pair of electrode area by equally spaced voltage or variant spaced voltage mode, it posits low-voltage on shorter isolated passage to make the mixture move to the direction of detector derived by higher field density; the position of low working voltage moves aroused by mixture moves so as to form moving gradient electric field at isolated passage to make substance finish isolation analysis with the driven field density of moving electric field. The electrophoresis chip to fulfill the method comprises array electrode, isolate pipe, sampling pipe, sample pool, waste liquid pool and buffer solution pool, wherein electrode array is posited on the floor or side board of micro-separator pipe.
Description
Technical field
The invention belongs to technical field of analytical instruments, particularly relate to a kind of implementation method and corresponding integrated electrophoresis chip system of low operating voltage electrophoresis.
Background technology
Micro Electro Mechanical System (Micro Electromechanical System, MEMS) appearance and developing rapidly, fundamentally change the design philosophy and the manufacture method of some scientific instrument system, for microminiaturized, integrated, intelligent, high performance-price ratio analysis and testing technology and instrument development provide technical foundation and new opportunity.Studies show that both at home and abroad that at present the electrophoresis chip technology progressively moves to maturity as a kind of core technology of emerging micro-total analysis system, and begun to bring impact even revolution to many fields of life science.Conventional chip electrophoresis mostly directly is applied to 1 kilovolt～several kilovoltages separating pipe head and the tail two ends, produces separation field intensity (Tang Yanghua, Zhou Zhaoying, Ye Xiongying etc. about 200～1000V/cm.The manufacturing of capillary electrophoresis chip.Micrometer-Nanometer Processing Technology.2001 (1): 62-66),, reduce the higher operating voltage of chip electrophoresis the development and the application of chip technology had great importance at the micro-total analysis technology.The electric field that utilizes alternating voltage to produce the certain rule variation on split tunnel carries out electrophoretic separation research (4M.S.Talalry, J.P.H.Burt, JA Tame, R.Pethig, Electro-manipulation and separation of cells using electricfields[J] .Appl.Phys., 1996, (29): 2198-2203.Employing electric field controls and isolated cell/＜applied physics 〉), and also report (5Manz A to some extent of the thought that the changing electric field that utilizes direct current synchronous circulation isolation technics to be produced is finished the separation detection process, Verpoorte E, Effrnhausc S, etal., Plannar chip technology for capillaryelectrophoresis[J], Anal.Chem., 1994,66 (1): 567_571.Planar chip technology in the Capillary Electrophoresis/＜analytical chemistry〉[U.S.]).The motional electric field that is produced by alternating voltage has been used for carrying out cell separation and two electrochondrias separates, and it can drive cell with the motion of 25-30 μ m/s speed, and the density of the electrod-array of generation driving ripple can reach 25000, and electrode is wide and all are 10 μ m.But adopt and on electrophoresis chip, produce a mobile gradient electric field, carry out the research and the technology of electrophoresis and do not see that as yet ripe report is arranged than low dc voltage.The motion gradient fields that system's low-work voltage that we propose separates forms and control method, by array electrode by rights impressed DC voltage guarantee the electric field intensity in zone, at this moment low one to two order of magnitude of the separation voltage that impressed voltage can be more original.And design and produce integrated electrophoresis chip of low-voltage and correspondent control circuits based on this.
Summary of the invention
The objective of the invention is to provide the implementation method of low operating voltage electrophoresis, and design and produce Low Voltage Electrophoresis Chip and correspondent control circuits based on this, realize that in the hope of being used to the compartment analysis of different biochemical systems detects.
The object of the present invention is achieved like this:
1, the implementation method of low-voltage electrophoresis
According to the electrophoretic separation principle, the realization that the low-work voltage of electrophoresis chip separates is under the prerequisite of needed separation electric field in guaranteeing detachment process, reduce the operating voltage that is applied on the split tunnel, promptly on the separation raceway groove, carry out and divide section, alternate cycles to apply voltage, thereby realize the low-work voltage of electrophoretic separation.
For this reason, the inventive method is: form the separate mesh electrode arrays at the differential of electrophoresis chip from pipeline equal intervals l a series of electrodes that distribute, guarantee that each Disengagement zone has an electrode at least, and the design correspondent peripheral circuit is controlled the break-make of each electrode and direct supply, thereby the control separation voltage is applied to the location and time on the split tunnel, utilize peripheral control circuit, adopt equidistantly to apply voltage system or become spacing and apply voltage system, be applied to successively in every pair of electrode zone of separating pipe adding low-voltage, at the separation initial stage, utilize electrod-array that low-voltage is applied on the short split tunnel, compounding substances is moved to the detecting device direction under higher field intensity drives; Along with the motion of material, low-voltage apply also migration thereupon of position, on split tunnel, just form the gradient electric field of motion, make material the Driving Field of motional electric field pretend with under finish compartment analysis.。The control mode that low-voltage is separated has two kinds: equidistantly apply voltage and apply voltage with the change spacing.
Equidistantly applying in the voltage system, the distance that each impressed voltage applies is a certain value, thus separated sample component under the effect of a constant field intensity, move all the time, along with the motion of material, low-voltage applies the position and also changes thereupon, makes potpourri realize separating under motional electric field drives.
Become spacing and apply voltage system, the motion that promptly changes with material changes the spacing that voltage applies, thereby separated material is moved under the field intensity effect that changes, increase the scope that low-voltage applies this moment on split tunnel, still move in the formed field strength range of low-voltage to guarantee fastest and the slowest material.So circulation just forms the variation field intensity of moving on split tunnel, make separated material finish the compartment analysis process under the field intensity effect of this motion change.
Because electric field separates is that alternating voltage ground is applied on the separating pipe, then the electric field region that applies in the process to be produced at voltage need have certain overlapping, thereby the assurance material particle moves under effect of electric field all the time.
In the voltage application mode that realizes low operating voltage electrophoresis, electrode arranging on the microchip pipeline also is crucial factor, can adopt sidewall to arrange that electrode and sole arrangement electrode dual mode come the gradient electric field of specific implementation motion.2, low operating voltage electrophoresis chip: the structure of low operating voltage electrophoresis chip comprises: array electrode, separating pipe, sample channel, buffer solution pond and waste liquid pool that sample cell that the sample channel two ends are connected to and waste liquid pool, separating pipe two ends are connected to.The structural parameters of chip are: electrode separation L is 200-1000 μ m, and electrode widths W e is 50-100 μ m, and duct width Wc is 20-100 μ m, and pipeline depth is 10-50 μ m, adopts Si
3N
4As insulation course, the THICKNESS CONTROL of insulation course is at 1-3 μ m.The cover plate of this chip can adopt bonding silicon chip, bonding glass or direct PDMS polymkeric substance cover plate.Array electrode is arranged in the sidewall of separating pipe or is arranged in the bottom surface.According to the structural parameters of design, adopt SOI MEMS process technology to carry out corresponding technological design, mask design and preparation processing.The sweep circuit of CMOS technology is adopted in system's control, realizes separating on the raceway groove and divides section, alternate cycles to apply voltage.Description of drawings
The method synoptic diagram that the low operating voltage electrophoresis of Fig. 1 electrophoresis chip is realized;
The electrod-array arrangement of Fig. 2 low operating voltage electrophoresis chip, wherein Fig. 2 (a) is that electrode Fig. 2 (b) both sides layout electrode is arranged in the bottom;
Fig. 3 is the structural representation of Low Voltage Electrophoresis Chip;
Fig. 4 is a low operating voltage electrophoresis chip drive circuit synoptic diagram.
Embodiment
Below in conjunction with accompanying drawing the implementation method of low operating voltage electrophoresis of the present invention and chip are done further and to be described in detail.
1. the implementation of low operating voltage electrophoresis:
The formation of motion gradient electric field relies on a series of array electrode to realize, to add low-voltage in the detachment process in turn is applied in every pair of electrode zone of separating pipe, and each Disengagement zone should at least also keep an electrode, when guaranteeing that adjacent twice electrode pair switches, the electric field region that is produced has certain overlapping, thereby guarantees that material particle moves under effect of electric field all the time.
Referring to Fig. 1, at the separation initial stage, utilize electrod-array that low-voltage is applied on the short split tunnel, compounding substances under driving, higher field intensity is moved to the detecting device direction; Along with the motion of material, low-voltage apply also migration thereupon of position, on split tunnel, just form the gradient electric field of motion, make material the Driving Field of motional electric field pretend with under finish the compartment analysis process.What a of Fig. 1, b, c, d showed is exactly certain process that voltage applies the position, E=V/L.
The array electrode in the microchannel of Low Voltage Electrophoresis Chip of Ti Chuing can have two kinds of distribution modes therefrom, the first directly evenly is arranged in duct bottom, see Fig. 2 (a), at this moment the width of pipeline can not produce obvious influence to the distribution of pipeline internal electric field, and the width of pipeline depth, electrode and spacing are the key factors that influences the distribution of pipeline internal electric field.It two is that array electrode evenly is arranged in the pipeline both sides, sees Fig. 2 (b), and at this moment the degree of depth of pipeline can not produce obvious influence to the distribution of electric field, and the width of duct width, electrode and spacing are the key factors that influences the distribution of pipeline internal electric field.
The microelectrode array and the structural parameters of microchannel: the electrode separation L of this patent design are 200-1000 μ m, and electrode widths W e is 50-100 μ m, and duct width Wc is 20-100 μ m, and pipeline depth is 10-50 μ m.This patent adopts Si
3N
4As insulation course, for suitable, the existence that can guarantee insulation course forms electric field intensity to differential in pipeline influence very little the THICKNESS CONTROL of insulation course at 1-3 μ m.
The control of low-voltage electrophoresis is adopted equidistantly to apply voltage or become spacing and is applied voltage and realize:
In equidistantly applying the voltage implementation, form the separate mesh electrode arrays at distribute a series of electrodes of capillary separation channels equal intervals l, and design the break-make that correspondent peripheral circuit is controlled each electrode and direct supply, thereby the control separation voltage is applied to the location and time on the split tunnel.If separation voltage applies scope and is always electrode number n * l, when separating initial stage t=0, in time Δ t, utilize control circuit and separate mesh electrode array, at the 1st of separate mesh electrode array, (n+1) apply voltage V between the individual electrode, drive sample component and move to positive dirction; After the material that moves the most slowly in the sample component passes through the position of the 2nd electrode, promptly after the time t=Δ t, voltage V is applied to the 2nd by control circuit, (n+2) between the individual electrode, cancel the 1st simultaneously, (n+1) voltage between the individual electrode; After the material that moves the most slowly in the sample component passes through the position of the 3rd electrode, be after the time t=2 Δ t, by control circuit voltage V is applied to the 3rd, (n+3) between the individual electrode, cancel the 2nd simultaneously, (n+2) voltage between the individual electrode, and the like, up to the detecting device of compounding substances, thereby finish whole detachment process by the raceway groove end.Along with applying of motional electric field, compounding substances separates certain distance gradually owing to the difference of mobility, still in the formed field strength range of low-voltage, move in order to ensure fastest and the slowest material, the distance that voltage is applied will satisfy: 1) in the control time of the electrode pair first time, satisfy the slowest material of speed and arrive the voltage range that circulation is next time applied; 2) electrode pair is in the control time and the last time, and the slow and the fastest material of speed must be in the span scope that voltage applies.
Becoming spacing applies in the voltage system, thereby with apply equidistantly that the voltage system difference is when compounding substances because mobility different and separate certain distance gradually and exceed the scope that voltage applies, increase the scope that low-voltage applies this moment on split tunnel, still move in the formed field strength range of low-voltage to guarantee fastest and the slowest material.At the separation initial stage, the scope that separation voltage applies is electrode number n * l, utilizes control circuit at separate mesh electrode array the 1st, (n+1) applies voltage V between the individual electrode, separates field intensity E
1=V/nl drives sample component and moves to positive dirction.When the material that moves the most slowly in the sample component by after the position of the 2nd electrode, i.e. time t=t
1After, at constant time t
1In the section, with constant electric field strength E
1Voltage V is applied to separate mesh electrode array the 2nd, (n+2) between the individual electrode, and the like, until with t
1And E
1Be the cycle index c of control time with the 1st stage of separating field intensity
1Finish, the length that increases voltage then and applied, entering the control time is t
2, the separation field intensity is E
2=V/ (n+1) W
Sp, cycle index is C
2The 2nd stage.Like this recursion, applies separation voltage to alternate cycles at segmentation on split tunnel, gradient, thereby forms the gradient electric field that moves, and finishes the separation of sample component.
Because the gradient electric field detachment process is alternating voltage ground to be applied to separate on the raceway groove, so apply at voltage that to need electric field to have in the process certain overlapping, make sample can be subjected to effect of electric field, finish the moved apart process.Also promptly require: must make the slowest separate substance of speed at control time section t each time
jInterior displacement can be at t to guarantee this material greater than separate mesh electrode array pitch l
jThe voltage application time in arrive the scope that voltage next time applies.Simultaneously, at t
jControl time when finishing, fastest separate substance can not exceed the scope that this voltage applies.Alternating voltage control circuit of the present invention can adopt existing ripe control circuit form, realizes as adopting control circuit as shown in Figure 4.It is by multi-electrode scan drive circuit B; Scan control unit C and scanning voltage controller D, A controls formation to electrophoresis chip.
2. the structure of Low Voltage Electrophoresis Chip:
Referring to Fig. 3, the structure of low operating voltage electrophoresis chip comprises: array electrode 1, separating pipe 2 and sample channel 3, buffer solution pond 6 and waste liquid pool 7 that sample cell 4 that sample channel 2 two ends are connected to and waste liquid pool 5, separating pipe 2 two ends are connected to.The cover plate of this chip can adopt bonding silicon chip, bonding glass or direct PDMS polymkeric substance cover plate.Be arranged in the sidewall of separating pipe or be arranged in the bottom surface referring to Fig. 2 (a) and Fig. 2 (b) array electrode.Below be a kind of manufacture craft of chip::
Preparation soi wafer---zinc oxide-silicon dioxide---photoetching isolated area---deep etching---oxidation 1000nm and polycrystalline backfill---leveling photoetching---oxidation---photoetching electrode---boron diffusion---oxidation---lithography fair lead---sputter sial---photoetching lead-in wire---alloy---passivation---TOPESIDE photoetching---dry etching SiO2---dry etching silicon.
Utilize conventional cmos technology to make the alternating voltage control circuit of low operating voltage electrophoresis chip differential array electrode on passage, to control position and the time that separation voltage applies on the split tunnel.
Referring to Fig. 3, the structural parameters of chip are: chip width W=24mm, sample cell and crossing are apart from 2L
s=6mm, buffer pool and crossing distance L
b=5mm, liquid storage tank pore radius R=3mm, separating pipe width W
c=0.08mm, detecting electrode and array electrode apart from x=1.8mm, array electrode spacing d
e=1mm.
Advantage of the present invention is: 1. the moving grade field that the low-work voltage of setting up separates forms and control method, can pass through Array electrode by rights impressed DC voltage guarantees regional electric-field intensity, and at this moment applied voltage can more conventional electrophoresis The separation voltage of system reduces by one to two order of magnitude; 2. adopt the low-voltage electricity of SOI MEMS process technology design processing The swimming chip, have that separation voltage is low, power consumption is little, highly sensitive, be easy to the superiority such as batch production; 3. that sets up thus is low The voltage chip system is easy to realize system's microminiaturization, portability and the automation of real meaning, can be widely used in biochemical inspection The various fields such as field operations detection of survey, detection of clinical, legal medical expert's criminal investigation detection, environmental science and modern national defense.
Claims (6)
1. the low operating voltage electrophoresis implementation method of electrophoresis chip, it is characterized in that: the differential at electrophoresis chip forms the separate mesh electrode array from a series of electrodes of pipeline equal intervals/distribution, guarantee that each Disengagement zone has an electrode at least, and control the break-make of each electrode and direct supply by correspondent peripheral circuit, thereby the control separation voltage is applied to the location and time on the split tunnel, utilize peripheral control circuit, adopt equidistantly to apply voltage system or become spacing and apply voltage system, be applied to successively in every pair of electrode zone of separating pipe adding low-voltage, at the separation initial stage, utilize electrod-array that low-voltage is applied on the short split tunnel, compounding substances is moved to the detecting device direction under higher field intensity drives; Along with the motion of material, low-work voltage apply also migration thereupon of position, on split tunnel, just form the gradient electric field of motion, make material the Driving Field of motional electric field pretend with under finish compartment analysis.
2. the low operating voltage electrophoresis implementation method of electrophoresis chip according to claim 1, it is characterized in that: described equidistantly to apply voltage system as follows: establish separation voltage and apply scope and be always electrode number n * 1, when separating initial stage t=0, in time Δ t, at the 1st of separate mesh electrode array, (n+1) apply voltage V between the individual electrode, drive sample component and move to positive dirction; After the material that moves the most slowly in the sample component passes through the position of the 2nd electrode, promptly after the time t=Δ t, voltage V is applied to the 2nd by control circuit, (n+2) between the individual electrode, cancel the 1st simultaneously, (n+1) voltage between the individual electrode; After the material that moves the most slowly in the sample component passes through the position of the 3rd electrode, be after the time t=2 Δ t, by control circuit voltage V is applied to the 3rd, (n+3) between the individual electrode, cancel the 2nd simultaneously, (n+2) voltage between the individual electrode, and the like, up to the detecting device of compounding substances, thereby finish whole detachment process by the raceway groove end; Wherein the distance that applies of voltage will satisfy: 1) in the control time of the electrode pair first time, satisfy the slowest material of speed and arrive the voltage range that circulation is next time applied; 2) electrode pair is in the control time and the last time, and the slow and the fastest material of speed must be in the span scope that voltage applies.
3. the low-voltage electrophoresis implementation method of electrophoresis chip according to claim 1, it is characterized in that: it is as follows that described change spacing applies voltage system: the separation initial stage, the scope that separation voltage applies is electrode number n * 1, utilize control circuit at separate mesh electrode array the 1st, (n+1) apply voltage V between the individual electrode, separate field intensity E
1=V/n1 drives sample component and moves to positive dirction; When the material that moves the most slowly in the sample component by after the position of the 2nd electrode, i.e. time t=t
1After, at constant time t
1In the section, with constant electric field strength E
1Voltage V is applied to separate mesh electrode array the 2nd, (n+2) between the individual electrode, and the like, until with t
1And E
1Be the cycle index c of control time with the 1st stage of separating field intensity
1Finish, the length that increases voltage then and applied, entering the control time is t
2, the separation field intensity is E
2=V/ (n+1) W
Sp, cycle index is c
2The 2nd stage, like this recursion, applies separation voltage to alternate cycles at segmentation on split tunnel, gradient, thereby forms the gradient electric field that moves, and finishes the separation of sample component.
4. the low operating voltage electrophoresis implementation method of electrophoresis chip according to claim 1 is characterized in that: arranging employing sidewall layout electrode and the sole arrangement electrode dual mode of electrode on the microchip pipeline.
5. realize the low operating voltage electrophoresis chip of the described method of claim 1, it has array electrode, separating pipe, sample channel, buffer solution pond and waste liquid pool that sample cell that the sample channel two ends are connected to and waste liquid pool, separating pipe two ends are connected to, the cover plate of this chip can adopt bonding silicon chip, bonding glass or direct PDMS polymkeric substance cover plate, and electrod-array is arranged in the sidewall of microchip pipeline or is arranged in the bottom surface.
6. low operating voltage electrophoresis chip according to claim 5, it is characterized in that: wherein microelectrode array and differential from the structural parameters of pipeline are: electrode separation L is 200-1000 μ m, electrode widths W e is 50-100 μ m, duct width Wc is 20-100 μ m, pipeline depth is 10-50 μ m, adopts Si
3N
4As insulation course, the THICKNESS CONTROL of insulation course is at 1-3 μ m.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101441194B (en) * | 2008-12-25 | 2012-12-26 | 重庆大学 | Low voltage chip electrophoresis multi-section circulation type voltage exerting control system and method |
| CN105457692A (en) * | 2016-01-05 | 2016-04-06 | 重庆大学 | Microfluidic separation device and method |
| CN105797792A (en) * | 2016-03-28 | 2016-07-27 | 南京理工大学 | Driving method for low-voltage medium liquid drops on digital microfluidic chip |
| CN111007134A (en) * | 2019-12-17 | 2020-04-14 | 京东方科技集团股份有限公司 | Bio-electrophoresis device, apparatus and control method |
-
2005
- 2005-03-29 CN CN 200510020641 patent/CN1699989A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101441194B (en) * | 2008-12-25 | 2012-12-26 | 重庆大学 | Low voltage chip electrophoresis multi-section circulation type voltage exerting control system and method |
| CN105457692A (en) * | 2016-01-05 | 2016-04-06 | 重庆大学 | Microfluidic separation device and method |
| CN105797792A (en) * | 2016-03-28 | 2016-07-27 | 南京理工大学 | Driving method for low-voltage medium liquid drops on digital microfluidic chip |
| CN105797792B (en) * | 2016-03-28 | 2017-12-12 | 南京理工大学 | A kind of low-voltage medium drop driving method on digital microcurrent-controlled chip |
| CN111007134A (en) * | 2019-12-17 | 2020-04-14 | 京东方科技集团股份有限公司 | Bio-electrophoresis device, apparatus and control method |
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