CN109158135A - A kind of high-performance on piece capillary electrophoresis system based on nanostructure - Google Patents
A kind of high-performance on piece capillary electrophoresis system based on nanostructure Download PDFInfo
- Publication number
- CN109158135A CN109158135A CN201811147769.0A CN201811147769A CN109158135A CN 109158135 A CN109158135 A CN 109158135A CN 201811147769 A CN201811147769 A CN 201811147769A CN 109158135 A CN109158135 A CN 109158135A
- Authority
- CN
- China
- Prior art keywords
- capillary electrophoresis
- performance
- nanostructure
- electrophoresis system
- channel
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D57/00—Separation, other than separation of solids, not fully covered by a single other group or subclass, e.g. B03C
- B01D57/02—Separation, other than separation of solids, not fully covered by a single other group or subclass, e.g. B03C by electrophoresis
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/502707—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by the manufacture of the container or its components
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/416—Systems
- G01N27/447—Systems using electrophoresis
- G01N27/44756—Apparatus specially adapted therefor
- G01N27/44791—Microapparatus
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/10—Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/08—Geometry, shape and general structure
- B01L2300/0832—Geometry, shape and general structure cylindrical, tube shaped
- B01L2300/0838—Capillaries
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/12—Specific details about materials
Abstract
The invention discloses a kind of high-performance on piece capillary electrophoresis systems based on nanostructure for belonging to biotechnology.The present invention includes three-dimensional manometer array, micro-fluidic chip, the three-dimensional manometer array is integrated in the detection zone of sheet glass, micro-fluidic chip is intersected vertically to obtain (cross channel) by the channel of two micron-scales, and four tops in channel respectively connect 1 injection port.Entire detection process is integrated on micro-fluidic chip by the present invention by capillary electrophoresis system, thus sample volume is few, reaction speed is fast and simple and convenient.Meanwhile by realizing fluorescence enhancement in detection zone integrated metal nanostructure, the detection sensitivity of chip is substantially increased.
Description
Technical field
The invention belongs to field of biotechnology, and in particular to a kind of high-performance on piece Capillary Electrophoresis based on nanostructure
System.
Background technique
Conventional capillary electrophoresis tube (capillary electrophoresis, CE) is to drive quartz with high-voltage dc
In capillary in sample solution bio-molecular separation to be measured technology, separation and analysis for protein and DNA have important
Meaning.However, conventional capillary electrophoresis tube takes a long time, needs large-scale instrument and professional operator and fluorescence signal is in life to be measured
Object molecule (such as amino acid, protein and DNA) concentration is very weak when lower, so that analysis speed and separative efficiency are relatively low.
Summary of the invention
Sensitivity for conventional capillary electrophoresis tube is low, time-consuming and it is complicated for operation the disadvantages of, the present invention proposes a kind of base
In the high-performance on piece capillary electrophoresis system of nanostructure.Biomolecule (such as amino may be implemented in the capillary electrophoresis system
Acid, protein and DNA etc.) quick, highly sensitive detection.
The present invention is using a kind of nanometer for being based on glancing angle deposition (oblique angle deposition, OAD) technology
Structure preparation process.OAD is a kind of physical gas-phase deposite method, and principle is to be made by the diffusion of shadow effect and adatom
With growing controllable three-dimensional manometer array (such as Fig. 1) in substrate.Nano-array (gold, silver and the silica that the present invention integrates
Deng) average diameter is 50nm-200nm, with a thickness of 100nm-3000nm, spacing is 50nm-500nm (such as Fig. 2).
Further, above-mentioned nanostructure is controllably integrated in by the present invention by depositing template (deposition mask)
Sheet glass certain area (such as Fig. 3).The region is the detection zone of capillary electrophoresis system, and metal nano array therein can lead to
Cross the detection sensitivity that biomolecule is greatly improved in enhancement effect of fluorescence.
Further, the present invention is using a kind of micro-fluidic chip (such as Fig. 4) with " cross " channel.The chip is by one long one
Short two channels intersect vertically to obtain, and each channel top connects 4 injection ports altogether.CE may be implemented in the chip, wherein short channel
Sample introduction is provided, long-channel interception sample segment simultaneously realizes separation and detection, and high-voltage dc provides power for whole system.
Further, there are two types of preparation methods for the chip.The first preparation method (such as Fig. 5): it prepares there is cross channel
Dimethyl silicone polymer (PDMS), and pass through plasma reaction (plasma) and the sheet glass carrier key for being integrated with nanostructure
Conjunction obtains complete chip.Second of preparation method: the sheet glass that there is cross channel and be integrated with nanostructure is prepared, and is passed through
Plasma is bonded to obtain complete chip with smooth PDMS.
Further, voltage needed for obtaining CE by Labview software programming program controls and reacts time delay command, and passes
Be defeated by can external control high voltage power supply.Thus the external environment system formed can provide suitable electricity to four injection ports of the chip
It presses and reacts delay.
A kind of high-performance on piece capillary electrophoresis system detection speed based on nanostructure of the present invention is fast, high sensitivity,
Easy to operate, operator only needs first to be full of each channel of CE chip and injection port with solution, then an injection port is added wherein
Sample to be detected, the biomolecule in the sample will realize electrophoresis point under the control of computer program and external high pressure power supply
From, and capture separation signal in the detection zone of integrated metal nanometer rods and analyze processing.
The utility model has the advantages that the high-performance on piece capillary electrophoresis system proposed by the present invention based on nanostructure realizes biology
Quick, the highly sensitive detection of molecule.Entire detection process is integrated on micro-fluidic chip by the capillary electrophoresis system, thus
Sample volume is few, reaction speed is fast and simple and convenient.Meanwhile by realizing fluorescence in detection zone integrated metal nanostructure
Enhancing, substantially increases the detection sensitivity of chip.
Detailed description of the invention
Fig. 1 is glancing angle deposition schematic diagram.
Fig. 2 is silver nanoparticle array SEM figure.
Fig. 3 is partial region integrated nanometer structure chart on sheet glass.
Fig. 4 is " cross " channel junction composition of CE chip.
Fig. 5 is the sheet glass compositing chip figure of " cross " channel PDMS and integrated Silver nanorod.
Specific embodiment
The present invention is described in detail with reference to the accompanying drawings and examples, but implementation of the invention is not limited only to this.
Embodiment 1
It is deposited on the glass sheet using glancing angle deposition silver nanostructured
Before the deposition, sheet glass is ultrasonically treated in acetone, isopropanol and deionization (DI) water, then passes through nitrogen
It is dry.It is first the Ti of 10nm with deposition angles θ=0 ° evaporation thickness to enhance the adhesiveness between Silver nanorod and sheet glass
Film layer.Then tool thickness is deposited with deposition angles θ=86 ° is respectively 300nm, 500nm, 1000nm, 2000nm and 3000nm
Silver nanorod.
Embodiment 2
Prepare CE chip of the device on PDMS
First by positive photoetching SU8 photoresist, certain thickness SU8 silicon wafer template is prepared on silicon wafer.By A/B=1/
10 a certain amount of PDMS solution vacuumizes bubble removing after being mixed evenly, then limpid PDMS mixed liquor is added in SU8 silicon wafer
Template surface simultaneously solidifies 1h at 80 DEG C.The full wafer PDMS and PDMS comprising single cross channel cuts with a knife of gently tearing is cut
It separates and punches.PDMS is cleaned, process is first to be cleaned by ultrasonic 5min with acetone, then be cleaned by ultrasonic 5min with isopropanol, then use DI
Water ultrasonic cleaning 5min simultaneously uses N2Air-blowing is dry.To clean with cross channel PDMS and the glass for being integrated with metal nano-rod
Piece beats 60s at 60W plasma and bonding together, obtains integrated nanometer structure C E core of the device on PDMS at once
Piece.
Embodiment 3
Prepare the biological detection chip of device on the glass sheet
It first passes through front lighting and carves the band device glass piece template for preparing certain thickness photoresist on the glass sheet.Again by the mould
Plate is immersed in BOE solution wet etching, or uses dry etching to the template, obtains having certain thickness " ten on sheet glass
Word " channel.Nanostructure is deposited by detection zone of the glancing angle deposition to the sheet glass again.By the glass with single cross channel
The cutting of glass piece is separated and is cleaned up.To the clean sheet glass with device and integrated nanometer structure and 4 sample holes are beaten
Smooth PDMS substrate beats 60s at 60W plasma and bonding together, obtains device on the glass sheet integrated and receives at once
Rice structure C E chip.
Embodiment 4
Two kinds of amino acid are separated using the CE chip
To the device in embodiment 2 on PDMS CE chip.Configuration concentration is four boron of 10mM first in DI water
Sour sodium (sodium tetraborate decahydrate) buffer simultaneously removes impurity by aperture for 0.22 μm of membrane filter.
Arginine (Arg) and glycine (Gly) are dissolved in buffer respectively, concentration is 1mM, and all with 100:1 (v/v) ratio
Dark is blended in the molten fluorescein -5- isothiocyanates (fluoresceine-5-isothiocyanate) in acetone of 1mM
In overnight, realize two kinds of amino acid of fluorescent marker.Before analysis, by the amino acid of two kinds of fluorescent markers with 1:1 (v/v) ratio
It mixes and is diluted to 10 μM.
For convenience of statistics, it is along label clockwise from the injection port of left end to CE chip, V2, V3 and V4.By 20 μ L
Buffer sequentially add the injection port (V1, V2, V4) of chip left end 3, wait 1min solution automatic sampling to right end sample introduction
The 20 μ L solution are added thereto again after mouthful (V3), realize entire channel full of buffer.It is drawn in injection port V2 with dust-free paper
Buffer and be added 20 μ L concentration be 10 μM of two kinds of fluorescent markers amino acid mixed liquor.By computer-controlled high voltage power supply four
A platinum electrode delivery outlet is fixed in four injection ports, is controlled by Labview first with V1:V2:V3:V4=150V:300V:
150V:0V reacts 20s and realizes sample introduction, then realizes Amino acid score with V1:V2:V3:V4=300V:150V:0V:150V reaction 2min
From, pass through fluorescence microscope capture fluorescence separate signal.
Claims (9)
1. a kind of high-performance on piece capillary electrophoresis system based on nanostructure, which is characterized in that including three-dimensional manometer array,
Micro-fluidic chip, the three-dimensional manometer array are integrated in the detection zone of sheet glass.
2. a kind of high-performance on piece capillary electrophoresis system based on nanostructure according to claim 1, feature exist
In the three-dimensional manometer array diameter is 50nm-200nm, with a thickness of 100nm-3000nm, spacing 50nm-500nm.
3. a kind of high-performance on piece capillary electrophoresis system based on nanostructure according to claim 1, feature exist
In the three-dimensional manometer array can be gold, silver or silica.
4. a kind of high-performance on piece capillary electrophoresis system based on nanostructure according to claim 1, feature exist
In the detection zone that the three-dimensional manometer array is integrated in sheet glass by depositing template, the three-dimensional manometer array passes through glimmering
The detection sensitivity of photo-enhancement effect raising biomolecule.
5. a kind of high-performance on piece capillary electrophoresis system based on nanostructure according to claim 1, feature exist
In, the micro-fluidic chip be the cross channel structure to be intersected vertically by the channel of two micron-scales, the four of channel
A top respectively connects 1 injection port.
6. a kind of high-performance on piece capillary electrophoresis system based on nanostructure according to claim 1, feature exist
In, CE may be implemented in the micro-fluidic chip, wherein short channel provide sample introduction, long-channel interception sample segment and realize separation with
Detection.
7. a kind of high-performance on piece capillary electrophoresis system based on nanostructure according to claim 1, feature exist
In the preparation method of the micro-fluidic chip is the dimethyl silicone polymer that preparation has cross channel, and passes through plasma
It reacts and is bonded to obtain complete chip with the sheet glass carrier for being integrated with nanostructure.
8. a kind of high-performance on piece capillary electrophoresis system based on nanostructure according to claim 1, feature exist
In the preparation method of the micro-fluidic chip is that preparation has cross channel and is integrated with the sheet glass of nanostructure, and passes through
Plasma is bonded to obtain complete chip with smooth PDMS.
9. the application of the high-performance on piece capillary electrophoresis system described in claim 1 based on nanostructure, which is characterized in that first
Each channel of micro-fluidic chip and injection port are full of with solution, then an injection port addition sample to be detected wherein, in sample
Biomolecule realize and be separated by electrophoresis under the control of computer program and external high pressure power supply, and capture separation letter in detection zone
Number and analyze processing.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811147769.0A CN109158135A (en) | 2018-09-29 | 2018-09-29 | A kind of high-performance on piece capillary electrophoresis system based on nanostructure |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811147769.0A CN109158135A (en) | 2018-09-29 | 2018-09-29 | A kind of high-performance on piece capillary electrophoresis system based on nanostructure |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN109158135A true CN109158135A (en) | 2019-01-08 |
Family
ID=64892858
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811147769.0A Pending CN109158135A (en) | 2018-09-29 | 2018-09-29 | A kind of high-performance on piece capillary electrophoresis system based on nanostructure |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109158135A (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1904604A (en) * | 2006-08-01 | 2007-01-31 | 浙江大学 | Microvolume sample lead in device used in capillary tube electrophoresis and use method thereof |
| EP1567845A4 (en) * | 2002-07-08 | 2007-07-18 | John S Foster | Method and apparatus for sorting biological cells with a mems device |
-
2018
- 2018-09-29 CN CN201811147769.0A patent/CN109158135A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1567845A4 (en) * | 2002-07-08 | 2007-07-18 | John S Foster | Method and apparatus for sorting biological cells with a mems device |
| CN1904604A (en) * | 2006-08-01 | 2007-01-31 | 浙江大学 | Microvolume sample lead in device used in capillary tube electrophoresis and use method thereof |
Non-Patent Citations (2)
| Title |
|---|
| CHENYU XIAO ,ZHEN CAO 等: "Microfluidic-based metal enhanced fluorescence for capillary electrophoresis by Ag nanorod arrays", 《NANOTECHNOLOGY》 * |
| 肖陈妤: "银纳米棒阵列的金属荧光增强及其在微型毛细管电泳技术中的应用", 《中国优秀硕士学位论文全文数据库信息科技辑》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US11045806B2 (en) | Integrated type microfluidic electrochemical biosensor system and method for rapid biochemical analysis | |
| CN105772118B (en) | The SERS micro-fluidic chips and preparation method of integrated enhancing substrate on ITO electro-conductive glass | |
| CN103894248B (en) | A kind of single cell analysis micro-fluidic chip and system and single cell analysis method | |
| CN102174369B (en) | Micro-slit-structure-based full PDMS (polydimethylsiloxane) micro-fluidic cell capturing chip and manufacturing method thereof | |
| CN103616427B (en) | A kind of micro-fluidic electrochemica biological sensor-based system simultaneously detecting for the dissimilar blood serum designated object of prostate cancer | |
| CN107699485A (en) | Microelectrode fluidic chip and adjustable parameter single cell electroporation device | |
| CN102725060A (en) | Flow path device and sample processing device including same | |
| CN102621214A (en) | Method for carrying out deceleration and monomolecular capture on nucleic acid molecule based on solid-state nano hole | |
| CN106513066A (en) | Three-dimensional porous graphene micro-fluidic chip and graphene attachment method thereof | |
| CN102590314A (en) | Method for decelerating nucleic acid molecules in solid nanopore | |
| CN103879953B (en) | A kind of blade coating patterning method based on wax | |
| CN112973650B (en) | Nano-micro composite ball and preparation method and application thereof | |
| CN102590302A (en) | Method for producing gold nanoarray ultramicroelectrode | |
| CN103509852B (en) | Detection method for biomolecular probe-calibrated specific sites of DNA based on nanopore device | |
| US11614395B2 (en) | Miniaturized flow cell and system for single-molecule nanoconfinement and imaging | |
| CN103316723A (en) | Microfluidic electrophoresis chip | |
| CN109142754A (en) | A kind of protein detecting chip and preparation method thereof based on micro-nano fluid | |
| CN109158135A (en) | A kind of high-performance on piece capillary electrophoresis system based on nanostructure | |
| CN104267200B (en) | Cancer cell based on water passage surface micron order lines detects micro-fluidic chip and preparation method | |
| Hu et al. | Solid‐State Quad‐Nanopore Array for High‐Resolution Single‐Molecule Analysis and Discrimination | |
| CN106492895B (en) | A kind of device and method preparing nanotip pipette | |
| WO2019080905A1 (en) | Raman activated droplet sorting system and method | |
| TWI291025B (en) | An integral micro-dialysis electrophoresis chip having on-line labeling function and the analysis method thereof | |
| CN104198255B (en) | One kind is based on microscopical cell point device for mark and microscope | |
| CN112570051A (en) | Microfluidic chip for sorting DNA, and apparatus and method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20190108 |