CN106732842B - For the micro-fluidic chip and preparation method thereof without mark high content screening - Google Patents
For the micro-fluidic chip and preparation method thereof without mark high content screening Download PDFInfo
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Abstract
The invention belongs to the technical fields of micro-fluidic chip, and in particular to a kind of for the micro-fluidic chip and preparation method thereof without mark high content screening；The technical issues of solution are as follows: cell can be captured, position by providing one kind, be able to carry out high intension, micro-fluidic chip of unmarked drug screening and preparation method thereof；The technical solution of use are as follows: for the micro-fluidic chip without mark high content screening, the micro-fluidic chip is equipped with detection cell and a plurality of fluid channel, the a plurality of fluid channel is connected with the detection cell, and it is not attached to lead to each other between a plurality of fluid channel, multiple structural units are equipped in the detection cell, the multiple structural unit forms array type structure, and the structural unit includes: at least two keeper electrode and at least a pair for positioning cell for growing the growth electrode of nanodendrites structure.
Description
Technical field
The invention belongs to the technical fields of micro-fluidic chip, and in particular to a kind of for without mark high content screening
Micro-fluidic chip and preparation method thereof.
Background technique
High content screening mainly keep eucaryotic cell structure and it is fully functional under the premise of, while detection be screened sample pair
The influence of cellular morphology, cell growth, signal path and cytotoxicity etc., and it is subject to comprehensive analysis.In traditional height
Containing screening is usually that cell culture is carried out on microwell plate, applies medicine irritation, and be aided with automation equipment and high-resolution optics
Test macro is detected.
Common optical detection is based on fluorescence microscope, may be implemented in conjunction with laser confocal scanning technology high-resolution
The imaging of unicellular cells device.But the technology needs to mark subcellular unit (nucleus, mitochondria) using fluorescent dye
Note, and the bio-toxicity of fluorescent dye seriously affects the long-time observation and research of cell, in addition, the fluorescence of non-specific binding
Dye molecule can introduce background noise, reduce detection sensitivity.Different from fluorescent method, SERS technology is not necessarily to dye molecule mark
Know, multiple molecule functional groups can be detected simultaneously, the structural information and drug to cell, drug are in the metabolic process
The variation that molecular structure is occurred carries out the research of unimolecule magnitude.In addition, micro-fluidic chip has life relative to microwell plate
Object reagent and the feature that cell dosage is few, Large-scale Screening is high-efficient are of great significance for reducing drug development cost.Cause
This, it is to carry out the effective way without mark high content screening that SERS technology is combined with microflow control technique.
Cui Yiping (62,2014, BIOSENSORS BIOELECTRONICS) uses drop micro-fluidic chip, obtains
The Surface enhanced Raman spectroscopy of rhodanate in human serum and saliva.Average value is collected optical signal as a whole, and experiment can
To obtain quantitative repeatability well.But this method is due to that cannot capture well cell, be positioned and training
It supports, still faces the problem of extracellular matrix environment variation, the cell model in drop lacks mutual between cell and matrix
Effect, this may block cell-signaling pathways, inhibit differential protein and related gene expression.
Summary of the invention
The present invention overcomes the shortcomings of the prior art, technical problem to be solved are as follows: providing one kind can be to cell
It captured, positioned, enough carry out label-free, high content screening micro-fluidic chip and preparation method thereof.
In order to solve the above-mentioned technical problem, the technical solution adopted by the present invention are as follows: for without mark high content screening
Micro-fluidic chip, the micro-fluidic chip be equipped with detection cell and a plurality of fluid channel, a plurality of fluid channel with the inspection
It surveys pond to be connected, and is not attached to lead to each other between a plurality of fluid channel, multiple structural units are equipped in the detection cell, it is the multiple
Structural unit forms array type structure, and the structural unit includes: at least two for positioning the keeper electrodes and at least of cell
A pair is for growing the growth electrode of nanodendrites structure.
Preferably, the structural unit is three annulus shapes, and three annulus are the different concentric loop of diameter, inside
Annulus is equipped at least two tips electrode；At least two circular electrodes are equipped on middle annulus, be with above-mentioned circular electrode etc.
Branch, middle annulus are divided into more equal portions, are equipped with point on each circular electrode；Multiple tip electricity are equipped in outer toroid
Pole, the circular electrode on point electrode and middle annulus in outer toroid correspond；Point electrode in the interior annular is two-by-two
Between form a pair of of growth electrode, the point electrode in the outer toroid and the point of the circular electrode on corresponding middle annulus
A pair of of growth electrode is formed, the non-point of the circular electrode is keeper electrode.
Preferably, the structural unit is double circular ring structures, and two annulus are the different concentric loop of diameter, inside
Annulus is equipped at least two tips electrode, and at least two circular electrodes are equipped in outer toroid, be with above-mentioned circular electrode etc.
Branch, outer toroid are divided into more equal portions, and the point electrode in the interior annular forms a pair of of growth electrode between any two, described
Circular electrode is keeper electrode.
Preferably, the structural unit is double circular ring structures, and two annulus are the different concentric loop of diameter, inside
Annulus is equipped at least two circular electrodes, and using above-mentioned circular electrode as Along ent, outer toroid is divided into more equal portions, Mei Yiyuan
It is equipped with point on shape electrode, multiple point electrodes are equipped in outer toroid, on the point electrode and interior annular in outer toroid
Circular electrode correspond；Point electrode in the outer toroid and the point shape of the circular electrode in corresponding interior annular
Electrode is grown in a pair, and the non-point of the circular electrode is keeper electrode.
Preferably, the micro-fluidic chip includes: the substrate being successively bonded together, substrate and transparent cover, the knot
Structure unit is set in the substrate, and the detection cell and a plurality of fluid channel are all set on the substrate, and the detection cell
Top cover be the transparent cover, the transparent cover be equipped with multiple solution entrances, the multiple solution entrance and institute
It states a plurality of fluid channel and corresponds connection.
Preferably, a plurality of fluid channel includes: cell liquid fluid channel, culture medium fluid channel, drug fluid channel, plating solution
Fluid channel, deionized water fluid channel and waste liquid fluid channel are equipped with one-way conduction valve in above-mentioned fluid channel.
Preferably, comprising the following steps: S10, using MEMS technology, production includes keeper electrode and growth electricity in substrate
The array architecture unit of pole makes detection cell and a plurality of fluid channel on substrate, multiple solution is made in transparent cover and are gone out
Substrate, substrate and transparent cover are successively bonded together by entrance；S20, the electrolyte containing noble metal is passed through into transparent cover
On a solution entrance, flow through fluid channel, be passed through in detection cell；S30, the indirect current between every two growth electrode,
To grow the nanodendrites structure that growth on the nearest tip of electrode distance forms noble metal at two；S40, noble metal nano branch
After crystal structure is formed, deionized water is passed through into detection cell, to rinse detection cell and fluid channel.
Preferably, in step S10, the utilization MEMS technology, production includes keeper electrode and growth electricity in substrate
The array architecture unit of pole, specifically includes: S101, depositing first layer metal layer in substrate and etches, forms growth electrode
Electrical cable；S102, the first layer insulating of deposition simultaneously etch, and expose electrode window through ray；S103, deposition second layer metal layer are simultaneously
Etching, forms the electrical cable of keeper electrode；S104, the second layer insulating of deposition simultaneously etch, and expose electrode window through ray；S105,
Deposition third layer metal layer simultaneously etches, and forms growth electrode and keeper electrode.
Preferably, it after completing a drug screening, between the growth electrode that every two growth has nanodendrites structure, applies
The alternating current of amplitude is increased, fuse nanodendrites structure, is passed through deionized water flushing.
Preferably, the electrolyte containing noble metal is solion or complex solution, and every a pair is grown between electrode
The amplitude of the alternating current of application is greater than 3V, and frequency values are between 1kHz and 10MHz.
Compared with the prior art, the invention has the following beneficial effects:
1, the multiple structural units being arranged in the detection cell on the micro-fluidic chip in the present invention, are in array type structure, can
To realize high-throughput drug screening.Structural unit includes keeper electrode for positioning cell and for growing nanodendrites knot
The growth electrode of structure.Using growth electrode, can on the position of cell capture prefabricated SERS substrate, realize cell and its drug
The SERS in situ detection of reaction.Using keeper electrode, cell can be captured, be positioned and adhere-wall culture.Finally realize
Unicellular, high intension, drug in conjunction with SERS without mark and automatically screening analysis.
2, the present invention in, after completing a drug screening, every two growth have nanodendrites structure growth electrode it
Between, apply the alternating current of amplitude, fuse nanodendrites structure, then passes to deionized water flushing, is restored to chip originally
The state of nanodendrites structure is not grown, to prepare for the next use, chip is made to reach reusable purpose.
Detailed description of the invention
The present invention will be further described in detail with reference to the accompanying drawing.
Fig. 1 is that the structure for the micro-fluidic chip without mark high content screening that the embodiment of the present invention one provides is shown
It is intended to；
Fig. 2 is used for structure list in the micro-fluidic chip without mark high content screening for what the embodiment of the present invention one provided
The structural schematic diagram of member；
Fig. 3 is provided by Embodiment 2 of the present invention for structure list in the micro-fluidic chip without mark high content screening
The structural schematic diagram of member；
Fig. 4 is used for structure list in the micro-fluidic chip without mark high content screening for what the embodiment of the present invention three provided
The structural schematic diagram of member；
Fig. 5 is used for electrode in the micro-fluidic chip without mark high content screening for what the embodiment of the present invention one provided
Line schematic diagram；
Fig. 6 to Figure 10 is used in the micro-fluidic chip without mark high content screening for what the embodiment of the present invention one provided
The fabrication processing figure of structural unit；
Figure 11 is the structural schematic diagram for the cell drug screening system that the embodiment of the present invention one provides；
In figure: 1 is detection cell, and 2 be fluid channel, and 3 be structural unit, and 4 be substrate, and 5 be substrate, and 6 be transparent cover, and 7 are
Solution entrance, 8 be external pad, and 9 be cell observing and controlling chip, and 10 be chip TT&C system, and 11 be Raman test macro, and 12 are
Fluorometric investigation system, 13 be liquid feeding system, and 31 be keeper electrode, and 32 make a living long electrode, and 41 be first layer metal layer, and 42 are
First layer insulating, 43 be second layer metal layer, and 44 be the second layer insulating, and 45 be third layer metal layer.
Specific embodiment
In order to make the object, technical scheme and advantages of the embodiment of the invention clearer, below in conjunction with the embodiment of the present invention
In attached drawing, technical scheme in the embodiment of the invention is clearly and completely described, it is clear that described embodiment is
A part of the embodiments of the present invention, instead of all the embodiments；Based on the embodiments of the present invention, ordinary skill people
Member's every other embodiment obtained without creative efforts, shall fall within the protection scope of the present invention.
Fig. 1 is that the structure for the micro-fluidic chip without mark high content screening that the embodiment of the present invention one provides is shown
It is intended to, as shown in Figure 1, the micro-fluidic chip is equipped with detection for the micro-fluidic chip without mark high content screening
Pond 1 and a plurality of fluid channel 2, a plurality of fluid channel 2 are connected with the detection cell 1, and between a plurality of fluid channel 2 each other not
It is connected, multiple structural units 3 is equipped in the detection cell 1, the multiple structural unit 3 forms array type structure, the knot
Structure unit 3 includes: at least two keeper electrode 31 and at least a pair for positioning cell for growing nanodendrites structure
Grow electrode 32.
When positioning cell, keeper electrode 31 adds traveling wave voltage, the phase difference between adjacent positioned electrode 31 are as follows: 360 degree/
All keeper electrode numbers, for example the quantity of keeper electrode 31 is two, then 180 degree is differed between adjacent positioned electrode 31,
The quantity of keeper electrode 31 is three, then 120 degree are differed between adjacent positioned electrode 31, if the quantity of keeper electrode 31 is six
It is a, then 60 degree are differed between adjacent positioned electrode 31.
Specifically, the micro-fluidic chip can include: substrate 4, substrate 5 and the transparent cover 6 being successively bonded together, institute
It states structural unit 3 to be set in the substrate 4, the detection cell 1 and a plurality of fluid channel 2 are all set on the substrate 5, and institute
The top cover for stating detection cell 1 is the transparent cover 6, and the transparent cover 6 is equipped with multiple solution entrances 7, the multiple molten
Liquid entrance 7 is connected to a plurality of one-to-one correspondence of fluid channel 2.
More specifically, the substrate 4 is insulating materials, or to be covered with the silicon materials of insulating layer thereon.
It more specifically, the substrate 5 is curing type polymer, or is thermoplastic polymer, or polymerize for solvent volatile-type
Object.
Specifically, a plurality of fluid channel 2 can include: cell liquid fluid channel, culture medium fluid channel, drug fluid channel,
Plating solution fluid channel, deionized water fluid channel and waste liquid fluid channel can be equipped with one-way conduction valve, i.e. waste liquid in above-mentioned fluid channel 2
Fluid channel is for unidirectionally flowing out, other fluid channels are for unidirectionally flowing into detection cell 1.
Fig. 2 is used for structure list in the micro-fluidic chip without mark high content screening for what the embodiment of the present invention one provided
The structural schematic diagram of member, as shown in Fig. 2, the structural unit 3 is double circular ring structures, two annulus are different same of diameter
Heart annulus is equipped at least two tips electrode in interior annular, is equipped at least two circular electrodes, in outer toroid with above-mentioned circle
Shape electrode is Along ent, and outer toroid is divided into more equal portions, and the point electrode in the interior annular forms a pair of raw between any two
Long electrode 32, the circular electrode are keeper electrode 31.
The quantity of the annulus, the quantity of the keeper electrode 31 and the growth electrode 32 and setting position can bases
Actual needs is set, and in the present embodiment, the quantity of the keeper electrode 31 and the growth electrode 32 is six.It is described
Substrate 5 is using glass, silicon, polymetylmethacrylate or dimethyl silicone polymer PSMS as substrate, and there are six miniflows for etching
Road 2 and a detection cell 1；In addition, when making structural unit 3, the corresponding one layer of electrode connecting line layer of each annulus, adjacent electrode
A layer insulating is provided between connecting line layer.
Fig. 5 is used for electrode in the micro-fluidic chip without mark high content screening for what the embodiment of the present invention one provided
Line schematic diagram, as shown in figure 5, the electrode connecting line layer of the structural unit 3 in the present embodiment is by lateral connecting line layer and longitudinal line
Layer is constituted, wherein lateral connecting line layer is for corresponding fixed with neighbouring detecting elements respectively by six keeper electrodes 31 of outer toroid
Position electrode 31 connects, and longitudinal connecting line layer is used for interior annular six growth electrodes 32 are corresponding with neighbouring detecting elements respectively
It grows electrode 32 to connect, one end of last connecting line is connected on the external pad 8 of chip edge.
The multiple structural units 3 being arranged in detection cell 1 on micro-fluidic chip in the present embodiment are in array type structure,
It may be implemented high-throughput drug screening, and structural unit 3 includes keeper electrode 31 for positioning cell and receives for growing
The growth electrode 32 of rice pine-tree structure can prefabricated SERS substrate, realization on the position of cell capture using growth electrode 32
Cell and its SERS in situ detection of drug reflection can capture cell using keeper electrode 31, be positioned and adherent training
It supports, finally realizes multi-mode, the automatically screening analysis of drug unicellular, high-throughput, in conjunction with SERS.
Fig. 3 is provided by Embodiment 2 of the present invention for structure list in the micro-fluidic chip without mark high content screening
The structural schematic diagram of member, as shown in figure 3, the difference between this embodiment and the first embodiment lies in, the structural unit 3 can be double annulus
Structure, two annulus are the different concentric loop of diameter, are equipped at least two circular electrodes, in interior annular with above-mentioned circle
Shape electrode is Along ent, and outer toroid is divided into more equal portions, is equipped with point on each circular electrode, is equipped in outer toroid
Multiple point electrodes, the circular electrode on point electrode and interior annular in outer toroid correspond；Point in the outer toroid
Termination electrode forms a pair of of growth electrode 32, the non-point of the circular electrode with the point of the circular electrode in corresponding interior annular
End is keeper electrode 31.
Fig. 4 is used for structure list in the micro-fluidic chip without mark high content screening for what the embodiment of the present invention three provided
The structural schematic diagram of member, as shown in figure 4, the present embodiment and the difference of embodiment one, two are, the structural unit 3 can be three
Annulus shape, three annulus are the different concentric loop of diameter, and at least two tips electrode is equipped in interior annular；In
Annulus is equipped at least two circular electrodes, and using above-mentioned circular electrode as Along ent, middle annulus is divided into more equal portions, Mei Yiyuan
Point is equipped on shape electrode；Multiple point electrodes are equipped in outer toroid, on the point electrode and middle annulus in outer toroid
Circular electrode correspond；Point electrode in the interior annular forms a pair of of growth electrode 32, the outer circle between any two
Point electrode on ring forms a pair of of growth electrode 32, the circular electric with the point of the circular electrode on corresponding middle annulus
The non-point of pole is keeper electrode 31.
For the specific structure of the structural unit 3 in embodiment one, two, three, can be designed according to actual needs,
In, the diameter of the annulus where keeper electrode 31 is designed according to the diameter for being detected cell, for growing nanometer branch
Two growth electrodes 32 of crystal structure apart from the distance between nearest tip ∈ (0 μm, 40 μm], and two above-mentioned tips are opposite
Setting.
It, can be in cell by the structural unit 3 of embodiment one after being positioned for a cell
Portion's structure and its drug response carry out preferable detection observation, can be to the periphery of cell by the structural unit 3 of embodiment two
Structure and its drug response carry out preferable detection observation, can be tied to the middle part of cell by the structural unit 3 of embodiment three
Structure and its drug response and peripheral structure and its drug response carry out preferable detection observation, can be by adjusting keeper electrode 31
With the setting position of growth electrode 32, detection observation is carried out come the different parts to cell.
Correspondingly, the production method for the micro-fluidic chip without mark high content screening, it may include following steps:
S10, using MEMS technology, production includes keeper electrode 31 and the array architecture for growing electrode 32 in substrate 4
Unit 3 makes detection cell 1 and a plurality of fluid channel 2 on substrate 5, multiple solution entrances 7 is made in transparent cover 6, by base
Bottom 4, substrate 5 and transparent cover 6 are successively bonded together.
S20, the electrolyte containing noble metal is passed through into a solution entrance 7 in transparent cover 6, flows through fluid channel 2,
It is passed through in detection cell 1.
S30, the indirect current between every a pair of of growth electrode 32, in two growth electrodes 32 on nearest tip
Growth forms the nanodendrites structure of noble metal.
After S40, noble metal nano pine-tree structure are formed, it is passed through deionized water into detection cell 1, to rinse 1 He of detection cell
Fluid channel 2.
Specifically, in embodiment one being in the structural unit 3 of bicyclic six electrode structure, in step S10, the benefit
With MEMS technology, production includes keeper electrode 31 and the array architecture unit 3 for growing electrode 32 in substrate 4, can specifically be wrapped
It includes:
S101, first layer metal layer 41 is deposited in substrate 4 and is etched, form the electrical cable of growth electrode 32, such as
Shown in Fig. 6.
Specifically, before carrying out step S101, first select quartz glass as substrate, respectively with acetone, alcohol, go from
Sub- water is cleaned, and is dried for standby；Then photoetching and lift-off technique is recycled to process first layer metal layer 41.
Firstly, spin coating and drying on the glass sheet, mask plate is used to carry out photoetching development；Then, sputtering is with a thickness of 30nm
Adhesion layer of the titanium (Ti) as sheet glass and metal, then the gold (Au) sputtered with a thickness of 200nm；Finally, wafer is placed on dress
There is 30min in the ultrasonic tank of acetone, complete lift-off, realizes the graphical of first layer metal layer 41.
S102, the first layer insulating 42 of deposition simultaneously etch, and expose electrode window through ray, as shown in Figure 7.
Specifically, use PECVD again in substrate growth thickness for the SiO of 300nm2Insulating layer, and using reticle into
Row exposure, and use hydrofluoric acid solution etching insulating layer, make its leakage electrode.
S103, deposition second layer metal layer 43 simultaneously etch, and form the electrical cable of keeper electrode 31, as shown in Figure 8.
Specifically, the processing and fabricating of second layer metal layer 43, this layer of metal layer are carried out using photoetching and lift-off technique
It is Ti/Au layers, with a thickness of (30nm/200nm), image dissolves transverse electrode lead.
S104, the second layer insulating 44 of deposition simultaneously etch, and expose electrode window through ray, as shown in Figure 9；
Specifically, the SiO of another layer of 300nm thickness is generated using the method for PECVD2Insulating layer is etched by wet etching
The electrode hole of connection keeper electrode 31 and growth electrode 32 out.
S105, deposition third layer metal layer 45 simultaneously etch, and form growth electrode 32 and keeper electrode 31, as shown in Figure 10.
Specifically, keeper electrode 31 and growth electrode 32, this Ti/Au thickness are processed using photoetching and lift-off technique
Degree is 30nm/300nm.
More specifically, making detection cell 1 in step S10 on substrate 5 and a plurality of fluid channel 2, used technique including
The technologies such as soft lithographic, method of molding, pressure sintering, LIGA technique and laser ablation process.
It tentatively completes after chip, also to carry out the encapsulation of chip, when encapsulation: it first designs, make PCB circuit board,
On be provided with and the one-to-one turmeric pad of chip and the metal being connect with external control devices row needle；Then by chip glue
It is bonded on PCB, is attached the pad on the pad and circuit board on chip using gold ball bonding technique, board metal
Row's needle passes through conducting wire again and connect with external control devices.
After complete chip manufacturing comes out, carries out the preparation of SERS substrate: the electrolyte containing noble metal is passed through detection
In pond 1, by control exchange and the amplitude of DC offset voltage and the frequency of alternating voltage, nanodendrites structure can control
Pattern, position and the direction of growth just will form the nanodendrites structure of noble metal on every a pair of of growth 32 tip of electrode；It is expensive
After metal nano pine-tree structure is formed, it is passed through deionized water in detection cell 1, to rinse detection cell 1 and fluid channel 2, in case after
Continuous chip works normally.
Specifically, the alternating current applied between growth electrode 32 can be sine voltage, square-wave voltage, triangular wave electricity
Pressure can also be bias voltage, and amplitude is greater than 3V, and frequency is between 1KHz and 10MHz；The electrolyte being passed through can be ion
Solution or complex solution, more specifically, can be AuCl3·HCl·3H2O, Pd(acetate)2, In(CH3COO)3、
HauCl4·3H2O、H2PtCl6·6H2O、K2PtCl4And Cu(NO3)2Equal solution.
When cellular localization, apply identical sinusoidal signal on relative positioning electrode 31, is applied on adjacent positioned electrode 32
Adding phase phase difference is the sinusoidal signal of (360 degree/all keeper electrode numbers), and cell is under the action of the dielectric power born to electric field
The middle lower position movement of field strength.
After completing a drug screening, between the growth electrode 32 that every two growth has nanodendrites structure, apply big
The alternating current of amplitude, fuse nanodendrites structure, is passed through deionized water flushing, is restored to chip and does not grow nanodendrites originally
The state of structure makes chip reach reusable purpose to prepare for the next use.
Correspondingly, Figure 11 is the structural schematic diagram for the cell drug screening system that the embodiment of the present invention one provides, such as Figure 11
It is shown, the cell drug screening system include: cell observing and controlling chip 9(i.e. the present invention in micro-fluidic chip), chip observing and controlling
System 10, Raman test macro 11, fluorometric investigation system 12 and liquid feeding system 13.Wherein, chip TT&C system 10 be by
Arbitrary waveform signal generator composition, mainly there is following functions: overpotential and cellular localization signal needed for prepared by SERS substrate
It generates；Raman test macro 11 is mainly used to realize for cardiovascular and cerebrovascular, tumour, virus, the major diseases such as neuropsychiatric
The drug screening of prevention and treatment and the raman spectra in situ detection of interaction；Fluorometric investigation system 12 is used in experimental stage pair
Cellular morphology is observed and is recorded in real time, and is compared with Raman test result, can after formally screen to drug
Without using fluorometric investigation system 12；Liquid feeding system 13 is mainly made of syringe pump and flow path, for realizing in microchip
Various fluids are controlled, at the same also may be implemented it is automatic, quantitatively inject corresponding reagent.
When work, SERS substrate is first prepared, is rinsed with deionized water；Then cell liquid, culture solution, drug are successively passed through；
Finally observation detection；After, fuse SERS, and rinses well；When needing to carry out the drug screening of a new round, then original
Micro-fluidic chip on prepare SERS substrate again, repeat above step.
The present invention realizes high-throughput, efficient cell inspection by the design of cleverly more fluid channels and big detection cell
Survey analysis and drug screening；It is realized by special round keeper electrode and tip-growth electrode design using electrochemical techniques
Unicellular fixation and SERS substrate are prepared in situ and detect；The independent connection mode of electrode allows the present invention to pass through outside
The control of the electrode of diversification and automation finally realizes high intension, is unmarked for cardiovascular and cerebrovascular, tumour, virus, nerve essence
The multi-mode of the major diseases protective agents such as refreshing system, automatically screening analysis.
Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention., rather than its limitations；To the greatest extent
Pipe present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that: its according to
So be possible to modify the technical solutions described in the foregoing embodiments, or to some or all of the technical features into
Row equivalent replacement；And these are modified or replaceed, various embodiments of the present invention technology that it does not separate the essence of the corresponding technical solution
The range of scheme.
Claims (10)
1. the micro-fluidic chip is equipped with detection cell (1) and more for the micro-fluidic chip without mark high content screening
Fluid channel (2), a plurality of fluid channel (2) are connected with the detection cell (1), and between a plurality of fluid channel (2) each other
It is not attached to lead to, it is characterised in that: be equipped with multiple structural units (3) in the detection cell (1), the multiple structural unit (3) group
At array type structure, the structural unit (3) include: at least two for position cell keeper electrodes (31) and at least a pair
For growing the growth electrode (32) of nanodendrites structure.
2. according to claim 1 for the micro-fluidic chip without mark high content screening, it is characterised in that: described
Structural unit (3) is three annulus shapes, and three annulus are the different concentric loop of diameter, and at least two are equipped in interior annular
A point electrode；At least two circular electrodes are equipped on middle annulus, using above-mentioned circular electrode as Along ent, middle annulus is divided equally
For more equal portions, point is equipped on each circular electrode；Multiple point electrodes, the tip in outer toroid are equipped in outer toroid
Circular electrode on electrode and middle annulus corresponds；Point electrode in the interior annular forms a pair of of growth electricity between any two
Pole (32), the point electrode in the outer toroid form a pair of of growth electricity with the point of the circular electrode on corresponding middle annulus
Pole (32), the non-point of the circular electrode are keeper electrode (31).
3. according to claim 1 for the micro-fluidic chip without mark high content screening, it is characterised in that: described
Structural unit (3) is double circular ring structures, and two annulus are the different concentric loop of diameter, and at least two are equipped in interior annular
A point electrode is equipped at least two circular electrodes in outer toroid, and using above-mentioned circular electrode as Along ent, outer toroid is divided equally
For more equal portions, the point electrode in the interior annular forms a pair of of growth electrode (32) between any two, and the circular electrode is
Keeper electrode (31).
4. according to claim 1 for the micro-fluidic chip without mark high content screening, it is characterised in that: described
Structural unit (3) is double circular ring structures, and two annulus are the different concentric loop of diameter, and at least two are equipped in interior annular
A circular electrode, using above-mentioned circular electrode as Along ent, outer toroid is divided into more equal portions, is equipped with point on each circular electrode
End is equipped with multiple point electrodes in outer toroid, and the circular electrode one on the point electrode and interior annular in outer toroid is a pair of
It answers；Point electrode in the outer toroid forms a pair of of growth electrode with the point of the circular electrode in corresponding interior annular
(32), the non-point of the circular electrode is keeper electrode (31).
5. according to claim 1 for the micro-fluidic chip without mark high content screening, it is characterised in that: described
Micro-fluidic chip includes: the substrate (4) being successively bonded together, substrate (5) and transparent cover (6), and the structural unit (3) sets
It is placed on the substrate (4), the detection cell (1) and a plurality of fluid channel (2) are all set on the substrate (5), and the inspection
The top cover for surveying pond (1) is the transparent cover (6), and the transparent cover (6) is equipped with multiple solution entrances (7), described more
A solution entrance (7) corresponds with a plurality of fluid channel (2) and is connected to.
6. according to claim 1 for the micro-fluidic chip without mark high content screening, it is characterised in that: described
A plurality of fluid channel (2) include: cell liquid fluid channel, culture medium fluid channel, drug fluid channel, plating solution fluid channel, deionized water
Fluid channel and waste liquid fluid channel, above-mentioned fluid channel (2) is interior to be equipped with one-way conduction valve.
7. the production side for the micro-fluidic chip without mark high content screening as described in any in claim 1 to 6
Method, it is characterised in that: the following steps are included:
S10, using MEMS technology, production includes the array knot of keeper electrode (31) and growth electrode (32) on substrate (4)
Structure unit (3) makes detection cell (1) and a plurality of fluid channel (2) on substrate (5), makes multiple solution on transparent cover (6)
Substrate (4), substrate (5) and transparent cover (6) are successively bonded together by entrance (7)；
S20, the electrolyte containing noble metal is passed through into a solution entrance (7) on transparent cover (6), flows through fluid channel
(2), it is passed through in detection cell (1)；
S30, the indirect current between every a pair of of growth electrode (32), to grow electrode (32) apart from nearest tip in every a pair
It is upper to grow the nanodendrites structure for forming noble metal；
After S40, noble metal nano pine-tree structure are formed, to being passed through deionized water in detection cell (1), with rinse detection cell (1) and
Fluid channel (2).
8. the production method according to claim 7 for the micro-fluidic chip without mark high content screening, special
Sign is: in step S10, the utilization MEMS technology, production includes keeper electrode (31) and growth electrode on substrate (4)
(32) array architecture unit (3), specifically includes:
S101, first layer metal layer (41) are deposited on substrate (4) and are etched, form the electrical cable of growth electrode (32)；
S102, deposition the first layer insulating (42) simultaneously etch, and expose electrode window through ray；
S103, deposition second layer metal layer (43) simultaneously etch, and form the electrical cable of keeper electrode (31)；
S104, deposition the second layer insulating (44) simultaneously etch, and expose electrode window through ray；
S105, deposition third layer metal layer (45) simultaneously etch, and form growth electrode (32) and keeper electrode (31).
9. the production method according to claim 7 for the micro-fluidic chip without mark high content screening, special
Sign is: after completing a drug screening, between the growth electrode (32) that every two growth has nanodendrites structure, applying big
The alternating current of amplitude, fuse nanodendrites structure, is passed through deionized water flushing.
10. the production method according to claim 7 for the micro-fluidic chip without mark high content screening, special
Sign is: the electrolyte containing noble metal is solion or complex solution, is applied between every a pair of of growth electrode (32)
Alternating current amplitude be greater than 3V, frequency values are between 1kHz and 10MHz.
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| CN101523212A (en) * | 2006-10-12 | 2009-09-02 | 皇家飞利浦电子股份有限公司 | Fast biosensor with reagent layer |
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