CN104880447B - A kind of ZnO NRs/Au films complex three-dimensional micro-array biochip substrate and preparation method thereof - Google Patents
A kind of ZnO NRs/Au films complex three-dimensional micro-array biochip substrate and preparation method thereof Download PDFInfo
- Publication number
- CN104880447B CN104880447B CN201510313994.7A CN201510313994A CN104880447B CN 104880447 B CN104880447 B CN 104880447B CN 201510313994 A CN201510313994 A CN 201510313994A CN 104880447 B CN104880447 B CN 104880447B
- Authority
- CN
- China
- Prior art keywords
- zno nrs
- films
- film
- dimensional micro
- substrate
- 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.)
- Expired - Fee Related
Links
Landscapes
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The present invention provides a kind of new ZnO NRs/Au films complex three-dimensional micro-array biochip substrate and preparation method thereof, belongs to biological technical field.This method is first in substrate surface deposition film;Then obtained film is activated;ZnO NRs are finally grown on the film after obtained activation, obtain new ZnO NRs/Au films complex three-dimensional micro-array biochip substrate.The present invention also provides the new ZnO NRs/Au films complex three-dimensional micro-array biochip substrate that above-mentioned preparation method obtains, obtained biochip substrate is subjected to functional modification, microarray is write, the signal of microarray is obtained, compared with the signal obtained in commercialized substrate.Test result indicates that:Compared with existing business slide, new ZnO NRs/Au films complex three-dimensional micro-array biochip substrate of the present invention can improve fluorescence signal intensity from decades of times to hundreds of times.
Description
Technical field
The present invention relates to biological technical field, and in particular to a kind of ZnO NRs/Au films complex three-dimensional micro-array biochip
Substrate and preparation method thereof.
Background technology
With the startup and implementation of the Human Genome Project, biochip technology obtains more and more extensive concern.The mankind
Genome plan is the essential step that the mankind are stepped by announcement itself secret, is after Manhattan project and Apollo Moon-landing Project
There is the research of epoch-making significance in human history afterwards.Biochip is current protein science, the important work of genomics research
Tool, is twentieth century end biotechnology the next item up breakthrough invention, and the development for biotech industry can match in excellence or beauty Henry ford
Great revolution of the auto production line organized to auto industry, or the revolutionary development similar to semiconductor for computer industry, extensively
For justice, biochip refers in glass, silicon chip, plastic or other material, utilizes the works such as microelectronics, micromechanics, photoelectricity, automation
Industry technology is made applied to biology, the product of chemical molecular, and its effective object can be gene, protein or cell, tissue
Deng.The confidence level for being mainly characterized by analysis and accuracy of biochip technology are high, and analyze speed is fast, used sample and examination
Agent is few, can handle a large amount of samples simultaneously.
In recent years, micro-array biochip (Microarray) is increasingly becoming the important component of biochip, turns into
It is a kind of by it is more and more extensive applied to gene, protein, carbohydrate, cell and other biological component analysis method, there is height
The features such as flux, automation, few sample consumption.Its basic thought is using mechanically or chemically in the solid substrate of activation
Method prepares the matrix arrangement of various biomolecule, and then various living matters are entered by intermolecular specificity interaction
Row analysis detection.
The making of micro-array chip is usually directed to the selection and preparation of chip base material, the surface modification of chip, chip
The steps such as the selection of biological labled material.Chip base material is the core of micro-array biochip, preferable chip list
Face should have the characteristics that size is accurate, smooth, homogeneous, fluorescence inertia, and usually used chip base material is that commercialization carries glass
The thin slice of piece, silicon chip and high polymer.For the micro-array chip that the biomolecule of fluorescence labeling is formed, the intensity of signal
It is closely related with the property of base material in itself.Sometimes for realize it is overdelicate detection, it is necessary to take some any special measures come
Improve the signal intensity on micro-array biochip surface.But commercialized glass and silicon chip are as base material at present, its
Body is to the signal of chip surface and does not have amplification.With going deep into for research, in some researchs in chip surface load
Some polymer, form spacial framework, and this substrate is referred to as three-dimensional macromolecule substrate, this kind of research at present by
Step starts commercial applications;In other research, someone forms Porous Silicon structures using silicon chip, substrate is had three in itself
The space structure of dimension, at present this kind of research do not use on a large scale, and this is due to be needed in experimentation by large-scale
Etching apparatus etc., virtually add the preparation cost of chip.The principle of the above method is all to utilize the ratio for increasing base material
Surface area improves the fixed amount of biomolecule, to reach the purpose of enhancing signal.Using other methods increase signal intensity still
It is rare.
Surface plasmon resonance (Surface Plasmon Resonance, SPR) is the phasmon ripple of metal surface
A kind of covibration caused by coupling incident light, is a kind of very unique optical characteristics of metal Nano structure.When resonating
When, incident light is absorbed by metal surface, produces the electromagnetic field of local enhancing in metal surface, it will to the fluorescence in suitable distance
The fluorescence signal of molecule is strengthened.Recent years, strengthening fluorescence using SPR fields has many important answer in bio-sensing field
With.
The content of the invention
It is an object of the invention to provide plant a kind of ZnO NRs/Au films complex three-dimensional micro-array biochip substrate and its system
Using ZnO NRs three-dimensional structure and nanoscale Au films phasmon occurs under light illumination for Preparation Method, the biochip substrate
Covibration, the electromagnetic field of enhancing is produced, the fluorescence combined jointly to biochip surface using SPR fields and three-dimensional ZnO NRs
The signal of molecule is strengthened, and is improved and be have detected sensitivity.
A kind of preparation method of ZnO NRs/Au films complex three-dimensional micro-array biochip substrate, this method include:
Step 1:In substrate surface deposition film, described membrane structure includes:Strengthen adsorption layer and surface phasmon
Excitation layer, described enhancing adsorption layer include 0.1-5.0nm Cr or Ti, and surface phasmon excitation layer includes 1-100nm's
Au films;
Step 2:The film that step 1 is obtained is activated;
Step 3:ZnO NRs are grown on film after the activation that step 2 obtains, obtain ZnO NRs/Au films compound three
Tie up micro-array biochip substrate.
Preferably, described substrate is glass substrate, silicon chip or high polymer thin slice.
Preferably, described Cr or Ti thickness is 1nm, and the thickness of described Au films is 50nm.
Preferably, the sedimentation rate of the enhancing adsorption layer is 0.1-0.2nm/s, sedimentation time 5-50s, surface etc. from
The sedimentation rate of excimer excitation layer is 0.1-0.2nm/s, sedimentation time 250-500s.
Preferably, the activation process of described step two is:The film that step 1 is obtained is immersed in KMnO4The aqueous solution
Middle activation 10-100min, then reacts in 30-80 DEG C of baking oven.
Preferably, described KMnO4The concentration of the aqueous solution is 1-100mM.
Preferably, ZnO NRs specific methods are grown on the film of described step three after activation is:Step 2 is lived
Film after change is put into the growth solution being made up of Zn salting liquids, ammoniacal liquor and monoethanolamine, raw in 60-90 DEG C of water bath
Long 1-4h.
Preferably, described Zn salting liquids include ZnCl2Solution, Zn (NO3)2·6H2O solution, ZnSO4·7H2O solution
Or Zn (CH3COO)2·2H2O solution.
Preferably, Zn in described Zn salting liquids2+Concentration is 10-500mM.
The present invention also provides the ZnO NRs/Au film complex three-dimensional micro-array biochip substrates that above-mentioned preparation method obtains.
Beneficial effects of the present invention
The present invention provides a kind of preparation method of ZnO NRs/Au films complex three-dimensional micro-array biochip substrate, this method
First there is plasmon resonance property substrate in substrate surface deposition film, formation;Then obtained film is activated;Most
ZnO NRs are grown on the film after obtained activation afterwards, obtain ZnO NRs/Au film complex three-dimensional micro-array biochip bases
Bottom.The preparation method is simple, raw material is easy to get, and the ZnO NRs on Au films surface can be easy to be removed without destroying by diluted acid in addition
Au films, the reuse of Au film substrates is realized, the production cost of chip can be substantially reduced.Obtained biochip substrate can be used for
In the fluoroscopic examination of protein microarray biochip.
The present invention also provides the ZnO NRs/Au film complex three-dimensional micro-array biochip substrates that above-mentioned preparation method obtains,
The biochip substrate has three-dimensional spacial framework, fluorescence signal can be increased by way of improving fixed amount
By force;On the other hand, the SPR characteristics of the nanoscale Au films below three-dimensional structure, the electromagnetic field pair of local enhancing is produced under light illumination
Fluorescence signal further enhances, and both reach increase fluorescence signal intensity jointly, can improve detection sensitivity.The life that will be obtained
Thing chip base carries out functional modification, writes microarray, the signal of microarray is obtained, with the letter obtained in commercialized substrate
Number it is compared.Test result indicates that:Compared with existing business slide, by controlling experiment parameter and changing experiment condition
ZnO NRs/Au films complex three-dimensional micro-array biochip substrate of the present invention can improve fluorescence signal intensity from tens of
Arrive hundreds of times again.
Brief description of the drawings
Fig. 1 is the SEM figures for the ZnO NRs/Au film complex three-dimensional micro-array biochip substrates that embodiment 1 obtains;
Fig. 2 is the SEM sectional views for the ZnO NRs/Au film complex three-dimensional micro-array biochip substrates that embodiment 1 obtains;
Fig. 3 is the XRD for the ZnO NRs/Au film complex three-dimensional micro-array biochip substrates that embodiment 1 obtains;
Fig. 4 is that the diffusing reflection for the ZnO NRs/Au film complex three-dimensional micro-array biochip substrates that embodiment 1 obtains is ultraviolet
Spectrogram;
Fig. 5 is the SEM figures for the ZnO NRs/Au film complex three-dimensional micro-array biochip substrates that embodiment 3 obtains;
Fig. 6 is the SEM sectional views for the ZnO NRs/Au film complex three-dimensional micro-array biochip substrates that embodiment 3 obtains;
Fig. 7 is the XRD for the ZnO NRs/Au film complex three-dimensional micro-array biochip substrates that embodiment 3 obtains;
Fig. 8 is that the diffusing reflection for the ZnO NRs/Au film complex three-dimensional micro-array biochip substrates that embodiment 3 obtains is ultraviolet
Spectrogram;
Fig. 9 is the SEM figures for the ZnO NRs/Au film complex three-dimensional micro-array biochip substrates that embodiment 4 obtains;
Figure 10 is the SEM sectional views for the ZnO NRs/Au film complex three-dimensional micro-array biochip substrates that embodiment 4 obtains;
Figure 11 is the XRD for the ZnO NRs/Au film complex three-dimensional micro-array biochip substrates that embodiment 4 obtains;
Figure 12 is that the diffusing reflection for the ZnO NRs/Au film complex three-dimensional micro-array biochip substrates that embodiment 4 obtains is ultraviolet
Spectrogram;
Figure 13 is before the ZnO NRs/Au film complex three-dimensional micro-array biochips matrix epoxyization that embodiment 1 obtains is modified
Contact angle afterwards characterizes;
Figure 14 is before the ZnO NRs/Au film complex three-dimensional micro-array biochips matrix epoxyization that embodiment 3 obtains is modified
Contact angle afterwards characterizes;
Figure 15 is before the ZnO NRs/Au film complex three-dimensional micro-array biochips matrix epoxyization that embodiment 4 obtains is modified
Contact angle afterwards characterizes;
Figure 16 is the fluorescence intensity block diagram that embodiment 5 writes Cy3-IgG microarrays in three kinds of different bases;
Figure 17 is the fluorescence micrograph that embodiment 5 writes Cy3-IgG microarrays in three kinds of different bases;
Figure 18 is the fluorescence intensity block diagram that embodiment 6 writes Cy3-IgG microarrays in three kinds of different bases;
Figure 19 is the fluorescence micrograph that embodiment 6 writes Cy3-IgG microarrays in three kinds of different bases;
Figure 20 is the fluorescence intensity block diagram that embodiment 7 writes Cy3-IgG microarrays in three kinds of different bases;
Figure 21 is the fluorescence micrograph that embodiment 7 writes Cy3-IgG microarrays in three kinds of different bases.
Embodiment
A kind of preparation method of ZnO NRs/Au films complex three-dimensional micro-array biochip substrate, this method include:
Step 1:In substrate surface deposition film, described membrane structure includes:Strengthen adsorption layer and surface phasmon
Excitation layer, described enhancing adsorption layer include 1-5nm Cr or Ti, and surface phasmon excitation layer includes 1-100nm Au films;
Step 2:The film that step 1 is obtained is activated;
Step 3:ZnO NRs are grown on film after the activation that step 2 obtains, obtain ZnO NRs/Au films compound three
Tie up micro-array biochip substrate.
According to the present invention, in substrate surface deposition film, the method for deposition film is not particularly limited, it is preferred to use electricity
Beamlet is deposited or the method deposition film of magnetron sputtering, preferably will deposition during using the method for electron beam evaporation plating or magnetron sputtering
Vacuum in equipment is extracted into 2 × 10-4-5×10-4Pa, described sedimentation rate and sedimentation time select according to the size of film thickness
Select, the sedimentation rate of Cr or Ti the enhancing adsorption layer is 0.1-0.2nm/s, and sedimentation time 5-50s, Au surface phasmon swashs
The sedimentation rate for sending out layer is 0.1-0.2nm/s, sedimentation time 250-500s.Described substrate is not particularly limited, and is preferably
Glass substrate, silicon chip or high polymer thin slice, substrate before the use, need to be cleaned, and specific cleaning method preferably includes:Will
Substrate piranha solution (concentrated sulfuric acid and 30% dioxygen water volume ratio 3:1 mixing) soaked overnight, organic pollution is removed, so
Afterwards with a large amount of deionized water rinsed cleans, it is cleaned by ultrasonic three times.Described membrane structure includes:Strengthen adsorption layer and be arranged on increasing
Surface phasmon excitation layer on strong adsorption layer, described enhancing adsorption layer include 0.1-5.0nm Cr or Ti, preferably Cr or
The thickness of Ti enhancing adsorption layers is 1nm, and enhancing adsorption layer is used to improve the tack between substrate and metal, surface phasmon
Excitation layer includes 1-100nm Au films, preferably 50nm Au films.
According to the present invention, film obtained above is activated, film is preferably immersed in KMnO4In the aqueous solution,
10-100min is activated at 50 DEG C, then reacts 10-60min in 30-80 DEG C of baking oven, so that priming reaction is fully carried out;
Described KMnO4The concentration of the aqueous solution is preferably 1-100mM.
According to the present invention, by the deionized water rinsed clean of the film after above-mentioned activation, then on film after activation
ZnO NRs are grown, specific method is preferably:Film after activation is put into by Zn salting liquids, 2-8% (V/V) ammoniacal liquor (28%
Wt) and in the growth solution of 5-20% (V/V) monoethanolamine (>=98%) composition, 1-4h is grown in 60-90 DEG C of water bath.
Described Zn salting liquids preferably include ZnCl2Solution, Zn (NO3)2·6H2O solution, ZnSO4·7H2O solution or Zn
(CH3COO)2·2H2O solution;Zn in described Zn salting liquids2+Concentration is preferably 10-500mM, after the completion of growth, uses deionization
Water rinsed clean, naturally dry, produce ZnO NRs/Au film complex three-dimensional micro-array biochip substrates.
The present invention also provides the ZnO NRs/Au film complex three-dimensional micro-array biochip substrates that above-mentioned preparation method obtains.
ZnO NRs/Au films complex three-dimensional micro-array biochip substrate obtained above is applied in protein microarray
In, specific method is:
Step 1:ZnO NRs/Au film complex three-dimensional micro-array biochip substrates are immersed in 1%-20%'s (V/V)
In 3- glycydoxies trimethoxy silane (GPTS) ethanol solution, 1-6h is incubated in the environment of 30-60 DEG C, so
Place into afterwards in 80-200 DEG C of vacuum drying oven, vacuumize and preserve 1-6h, it is modified biochip substrate surface, shape
Viability base group modification layer, surface binding biomolecules form specific micro-array biochip, obtain the chip of epoxy-modified;
Step 2:The chip of the epoxy-modified obtained to step 1 is micro- using contact spotting robot writing protein
Array, the sampling liquid used in experiment are 0.01M PBS solutions (PH=7.4), NMF glycerine (volume fraction 1%-30%),
Surfactant Triton X-100 (volume fraction 0.002%-0.008%) and 10-100 μ g/ml Cy3-IgG mixing
Liquid;Chip cultivates 6-15h in relative humidity 30-80%, the permanent steady wet tank of 30-40 DEG C of temperature after point sample, is used after cultivation
0.05M TBS solution is cleaned 2-6 times, each 3-10min, and deionized water is cleaned 2-6 times, each 3-10min, N after cleaning2Blow
It is dry, obtain the micro-array chip containing Cy3-IgG;
Step 3:Using the microarray scanner with 532nm lasers step 2 is obtained containing the micro- of Cy3-IgG
Array chip chip is scanned, and specific sweep parameter is:Resolution ratio:5–40μm;Laser power:60-90%;Photomultiplier transit
Coefficient:400-800, obtain microarray data.
For a further understanding of the particular content of the present invention, the present invention is carried out with reference to specific embodiment further
Describe in detail.
Zn (NO are used on the 0nm Au films of comparative example 13)2·6H2O grows ZnO NRs as Fluorescence Increasing base as Zn salt
Bottom
Glass substrate is used into piranha solution (concentrated sulfuric acid and 30% dioxygen water volume ratio 3:1 mixing) soaked overnight, remove
The organic pollutions such as degreasing, then glass substrate is arranged on rack for cleaning, it is clear with a large amount of deionized water rinsed cleans, ultrasound
Wash three times, 20min/ times, after thoroughly cleaning, be put into growth solution, 2h, described growth solution are reacted in 75 DEG C of water-bath
Composition:100mM Zn (NO3)2·6H2The O aqueous solution, 4% (V/V) ammoniacal liquor (28%wt) and 10% (V/V) monoethanolamine (>=
98%), rinsed after growth with deionized water, naturally dry, produce ZnO NRs micro-array biochip substrates.
Embodiment 1 uses Zn (NO on 50nm Au films3)2·6H2O grows ZnO NRs as Fluorescence Increasing as Zn salt
Substrate
Step 1:The deposition of Au films on glass substrate
Glass substrate is used into piranha solution (concentrated sulfuric acid and 30% dioxygen water volume ratio 3:1 mixing) soaked overnight, remove
The organic pollutions such as degreasing, then glass substrate is arranged on rack for cleaning, it is clear with a large amount of deionized water rinsed cleans, ultrasound
Wash three times, 20min/ times, after thoroughly cleaning, thin film deposition is carried out to glass substrate surface using electron-beam coating equipment, will be heavy
Vacuum in product equipment is extracted into 2 × 10-4, described membrane structure includes:Strengthen adsorption layer 1nm Cr films and surface phasmon
Excitation layer 50nm Au films, described deposition Cr films sedimentation time is 20s, sedimentation rate 0.2nm/s;When depositing Au films deposition
Between be 500s, sedimentation rate 0.1nm/s;
Step 2:Au films are activated
Au films obtained above are immersed in 5mM KMnO4In, 30min is activated in the environment of 50 DEG C, then at 50 DEG C
30min is reacted in baking oven, so that priming reaction is fully carried out;
Step 3:ZnO NRs are grown on 50nm Au films
Above-mentioned sample after activation is taken out, with deionized water rinsed clean, it is molten that the sample after activation is then put into growth
In liquid, 2h, the composition of described growth solution are reacted in 75 DEG C of water-bath:100mM Zn (NO3)2·6H2The O aqueous solution, 4%
(V/V) ammoniacal liquor (28%wt) and 10% (V/V) monoethanolamine (>=98%), are rinsed with deionized water after growth, naturally dry, produced
ZnO NRs/Au film complex three-dimensional micro-array biochip substrates.
Fig. 1 is the SEM figures for the ZnO NRs/Au film complex three-dimensional micro-array biochip substrates that embodiment 1 obtains;Fig. 2 is
The SEM sectional views for the ZnO NRs/Au film complex three-dimensional micro-array biochip substrates that embodiment 1 obtains;Can be with from Fig. 1 and Fig. 2
Find out, for the diameter for the ZnO NRs that embodiment 1 obtains probably in 200-300nm, the length of ear of maize is about 4 μm, is perfect six side
The ZnO club shaped structures of crystallographic system.Fig. 3 is the ZnO NRs/Au film complex three-dimensional micro-array biochip substrates that embodiment 1 obtains
XRD, Fig. 3 can illustrate there is very strong diffraction maximum at 34.5 °, and it is the bar-shaped knot grown along (002) direction to illustrate material
Structure.Fig. 4 is the diffusing reflection ultraviolet spectra for the ZnO NRs/Au film complex three-dimensional micro-array biochip substrates that embodiment 1 obtains
Figure, Fig. 4 can be seen that ZnO NRs and be grown on 50nmAu films has obvious absorption in 300-600nm, and this is mainly due to
The phasmon effect of Au nano thin-films, Comparatively speaking, ZnO nanorod direct growth is on the glass sheet (equivalent to being grown in 0nm
Au films on), only have absorption at 300-350nm, and absorption intensity is little.This just illustrates that ZnO NRs are grown in
This substrate of 50nmAu films may more utilize light, so as to strengthen the fluorescence signal on chip base surface.
Embodiment 2 uses Zn (NO on 100nm Au films3)2·6H2O grows ZnO NRs as Fluorescence Increasing as Zn salt
Substrate
The step of with embodiment 1, is identical, and difference is, the thickness of Au films is 100nm.
Embodiment 3 uses ZnSO on 50nm Au films4·7H2O grows ZnO NRs as Fluorescence Increasing substrate as Zn salt
The step of with embodiment 1, is identical, and difference is, the Zn salting liquids in growth solution are ZnSO4·7H2O。
The SEM figures of the ZnO NRs/Au film complex three-dimensional micro-array biochip substrates that embodiment 3 obtains, SEM sectional views,
XRD and diffusing reflection ultraviolet spectrogram are shown in Fig. 5,6,7,8.
Embodiment 4 uses Zn (CH on 50nm Au films3COO)2·2H2O increases as Zn salt growth ZnO NRs as fluorescence
Strong basis bottom
The step of with embodiment 1, is identical, and difference is, the Zn salting liquids in growth solution are Zn (CH3COO)2·
2H2O。
The SEM figures of the ZnO NRs/Au film complex three-dimensional micro-array biochip substrates that embodiment 4 obtains, SEM sectional views,
XRD and diffusing reflection ultraviolet spectrogram are shown in Fig. 9,10,11,12.
Application of the embodiment 5ZnO NRs/Au film complex three-dimensional micro-array biochip substrates in protein microarray
Step 1:The ZnO NRs/Au film complex three-dimensional micro-array biochip substrates that embodiment 1 obtains are immersed in 5%
(V/V) in 3- glycydoxies trimethoxy silane (GPTS) ethanol solution, 3h is incubated in the environment of 40 DEG C,
Then place into 110 DEG C of vacuum drying oven, vacuumize and preserve 3h, it is modified biochip substrate surface, formed
Active group decorative layer, surface binding biomolecules form specific micro-array biochip, obtain the chip of epoxy-modified;
Figure 13 is before the ZnO NRs/Au film complex three-dimensional micro-array biochips matrix epoxyization that embodiment 1 obtains is modified
Contact angle afterwards characterizes, as can be seen from Figure 13, hydrophilic before modification after figure b is modification before figure a is modification, bright after modification
Aobvious hydrophobic, contact angle, which characterizes, significant change, illustrates that by successfully modification the operation of next step microarray writing can be carried out.
Step 2:The chip of the epoxy-modified obtained to step 1 is micro- using contact spotting robot writing protein
Array, the sampling liquid used in experiment are 0.01M PBS solutions (PH=7.4), 2.5% glycerine NMF glycerine, 0.004% table
Face activating agent Triton X-100 and 50 μ g/ml Cy3-IgG mixed liquor;Chip is in relative humidity 60%, temperature after point sample
8h is cultivated in 37 DEG C of permanent steady wet tank, is cleaned 3 times, each 5min using 0.05M TBS solution after cultivation, deionized water is clear
Wash 3 times, each 5min, N after cleaning2Drying, obtains the micro-array chip containing Cy3-IgG;
Step 3:Using the microarray scanner with 532nm lasers step 2 is obtained containing the micro- of Cy3-IgG
Array chip chip is scanned, and specific sweep parameter is:Resolution ratio:5μm;Laser power:70%;Photomultiplier transit coefficient:
550, obtain microarray data.
Figure 16 is that embodiment 5 (uses Zn (NO in three kinds of different bases on sheet glass, sheet glass3)2·6H2O grows ZnO
Zn (NO are used on NRs, 50nmAu film3)2·6H2O growths ZnO NRs) writing Cy3-IgG microarrays fluorescence intensity block diagram;
Figure 17 is that embodiment 5 (uses Zn (NO in three kinds of different bases on sheet glass, sheet glass3)2·6H2O growths ZnO NRs,
Zn (NO are used on 50nmAu films3)2·6H2O growths ZnO NRs) writing Cy3-IgG microarrays fluorescence micrograph, wherein scheming
B is the fluorescence micrograph for writing Cy3-IgG microarrays on the glass substrate, and it is that Zn (NO are used on glass that figure c, which is,3)2·
6H2The fluorescence micrograph of Cy3-IgG microarrays is write in O growth ZnO NRs substrate;Figure d is to be used on 50nm Au films
Zn(NO3)2·6H2The fluorescence micrograph of Cy3-IgG microarrays is write in O growth ZnO NRs substrate, can from Figure 16 and 17
To find out, fluorescence intensity is different in three kinds of different substrates, and it is flat glass substrate to grow ZnO NRs substrates on the glass sheet
25 times or so, and more than the 100 times of ZnO NRs/Au that fluorescence intensity in ZnO NRs substrates is flat glass substrate is grown on Au films
Film has obvious Fluorescence Increasing effect.
Application of the embodiment 6ZnO NRs/Au film complex three-dimensional micro-array biochip substrates in protein microarray
Identical with the step of embodiment 5, difference is, ZnO NRs/Au film complex three-dimensional micro-array bio cores used
Piece is what embodiment 3 was prepared.
Figure 14 is before the ZnO NRs/Au film complex three-dimensional micro-array biochips matrix epoxyization that embodiment 3 obtains is modified
Contact angle afterwards characterizes, before figure a is modification, after figure b is modification, it is seen from figure 14 that it is hydrophilic before modification, it is bright after modification
Aobvious hydrophobic, contact angle, which characterizes, significant change, illustrates that by successfully modification the operation of next step microarray writing can be carried out.
Figure 18 is that embodiment 6 (uses ZnSO in three kinds of different bases on sheet glass, sheet glass4·7H2O grows ZnO
ZnSO is used on NRs, 50nmAu film4·7H2O growths ZnO NRs) writing Cy3-IgG microarrays fluorescence intensity block diagram;Figure
19 be that embodiment 6 (uses ZnSO in three kinds of different bases on sheet glass, sheet glass4·7H2O growths ZnO NRs, 50nmAu films
It is upper to use ZnSO4·7H2O growth ZnO NRs) fluorescence micrograph of Cy3-IgG microarrays is write, wherein it is in glass base to scheme b
The fluorescence micrograph of Cy3-IgG microarrays is write on bottom, it is that ZnSO is used on glass that figure c, which is,4·7H2O growth ZnO NRs
Substrate on write Cy3-IgG microarrays fluorescence micrograph;Figure d is that ZnSO is used on 50nm Au films4·7H2O grows
The fluorescence micrograph of Cy3-IgG microarrays is write in ZnO NRs substrate, it is from Figure 18 and 19 as can be seen that raw on Au films
Fluorescence intensity is that more than 100 times of ZnO NRs/Au film of flat glass substrate has obvious Fluorescence Increasing in long ZnO NRs substrates
Effect.
Application of the embodiment 7ZnO NRs/Au film complex three-dimensional micro-array biochip substrates in protein microarray
Identical with the step of embodiment 5, difference is, ZnO NRs/Au film complex three-dimensional micro-array bio cores used
Piece is what embodiment 4 was prepared.
Figure 15 is before the ZnO NRs/Au film complex three-dimensional micro-array biochips matrix epoxyization that embodiment 4 obtains is modified
Contact angle afterwards characterizes, before figure a is modification, after figure b is modification, it can be seen from fig. 15 that it is hydrophilic before modification, it is bright after modification
Aobvious hydrophobic, contact angle, which characterizes, significant change, illustrates that by successfully modification the operation of next step microarray writing can be carried out.
Figure 20 is that embodiment 7 (uses Zn (CH in three kinds of different bases on sheet glass, sheet glass3COO)2·2H2O grows
Zn (CH are used on ZnO NRs, 50nmAu films3COO)2·2H2O growths ZnO NRs) writing Cy3-IgG microarrays fluorescence intensity
Block diagram;Figure 21 is that embodiment 7 (uses Zn (CH in three kinds of different bases on sheet glass, sheet glass3COO)2·2H2O grows
Zn (CH are used on ZnO NRs, 50nmAu films3COO)2·2H2O growths ZnO NRs) writing Cy3-IgG microarrays fluorescence microscopy
Photo, wherein figure b is the fluorescence micrograph for writing Cy3-IgG microarrays on the glass substrate, figure c, which is, to be used on glass
Zn(CH3COO)2·2H2The fluorescence micrograph of Cy3-IgG microarrays is write in O growth ZnO NRs substrate;Scheme d be
Zn (CH are used on 50nm Au films3COO)2·2H2The fluorescence microscopy of Cy3-IgG microarrays is write in O growth ZnO NRs substrate
Photo, from Figure 20 and 21 as can be seen that it is the 100 of flat glass substrate that fluorescence intensity in ZnO NRs substrates is grown on Au films
More times of ZnO NRs/Au films have obvious Fluorescence Increasing effect.
A kind of ZnO nano-rod array/Au film complex three-dimensional microarray substrates and its preparation are disclosed in the embodiment of the present invention
Method.Those skilled in the art can use for reference present disclosure, be suitably modified technological parameter realization.Pay particular attention to
It any similar replacement and change, is it will be apparent that they are all recognized to this area and those skilled in the relevant art to be
To be included within the present invention.The product and method of the present invention is described by preferred embodiment, it is evident that phase
Pass personnel are not departing from present invention, method described herein are being modified in spirit and scope or suitably change and combine
To realize invention application technology.Test result indicates that compared with existing business slide, by controlling experiment parameter and change
Experiment condition ZnO NRs/Au three-dimensional substrates of the present invention can improve fluorescence signal intensity from decades of times to hundreds of times.
Claims (5)
- A kind of 1. preparation method of ZnO NRs/Au films complex three-dimensional micro-array biochip substrate, it is characterised in that this method Including:Step 1:In substrate surface deposition film, described membrane structure includes:Enhancing adsorption layer and surface phasmon excite Layer, described enhancing adsorption layer include 0.1-5.0nm Cr or Ti, and surface phasmon excitation layer includes 1-100nm Au films;Step 2:The film that step 1 is obtained is activated:The film that step 1 is obtained is immersed in KMnO4It is living in the aqueous solution Change 10-100min, then reacted in 30-80 DEG C of baking oven;Described KMnO4The concentration of the aqueous solution is 1-100mM;Step 3:ZnO NRs are grown on film after the activation that step 2 obtains, specific method is:After step 2 is activated Film be put into the growth solution being made up of Zn salting liquids, ammoniacal liquor and monoethanolamine, grow 1- in 60-90 DEG C of water bath 4h;Obtain ZnO NRs/Au film complex three-dimensional micro-array biochip substrates;Described Zn salting liquids include ZnCl2Solution, Zn (NO3)2·6H2O solution, ZnSO4·7H2O solution or Zn (CH3COO)2·2H2O solution;Zn in described Zn salting liquids2+Concentration is 10-500mM.
- A kind of 2. preparation side of ZnO NRs/Au films complex three-dimensional micro-array biochip substrate according to claim 1 Method, it is characterised in that described substrate is glass substrate, silicon chip or high polymer thin slice.
- A kind of 3. preparation side of ZnO NRs/Au films complex three-dimensional micro-array biochip substrate according to claim 1 Method, it is characterised in that described Cr or Ti thickness are 1nm, and the thickness of described Au films is 50nm.
- A kind of 4. preparation side of ZnO NRs/Au films complex three-dimensional micro-array biochip substrate according to claim 1 Method, it is characterised in that the sedimentation rate of the enhancing adsorption layer be 0.1-0.2nm/s, sedimentation time 5-50s, and surface etc. is from sharp The sedimentation rate of first excitation layer is 0.1-0.2nm/s, sedimentation time 250-500s.
- 5. the ZnO NRs/Au film complex three-dimensional micro-array bio cores that the preparation method described in claim 1-4 any one obtains Piece substrate.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201510313994.7A CN104880447B (en) | 2015-06-09 | 2015-06-09 | A kind of ZnO NRs/Au films complex three-dimensional micro-array biochip substrate and preparation method thereof |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201510313994.7A CN104880447B (en) | 2015-06-09 | 2015-06-09 | A kind of ZnO NRs/Au films complex three-dimensional micro-array biochip substrate and preparation method thereof |
Publications (2)
Publication Number | Publication Date |
---|---|
CN104880447A CN104880447A (en) | 2015-09-02 |
CN104880447B true CN104880447B (en) | 2018-02-13 |
Family
ID=53948023
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201510313994.7A Expired - Fee Related CN104880447B (en) | 2015-06-09 | 2015-06-09 | A kind of ZnO NRs/Au films complex three-dimensional micro-array biochip substrate and preparation method thereof |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN104880447B (en) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN112611861B (en) * | 2020-11-23 | 2024-03-29 | 武汉世纪康敏生物科技有限公司 | Fluorescent immunodetection chip and preparation method thereof |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102901715A (en) * | 2012-11-07 | 2013-01-30 | 吉林大学 | Fluorescence enhanced microarray biochip based on micro/nano periodic structures and method for preparing same |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103147133B (en) * | 2013-02-25 | 2015-03-25 | 西南大学 | Three-dimensional carrier of microarray biochip and preparation method thereof |
-
2015
- 2015-06-09 CN CN201510313994.7A patent/CN104880447B/en not_active Expired - Fee Related
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102901715A (en) * | 2012-11-07 | 2013-01-30 | 吉林大学 | Fluorescence enhanced microarray biochip based on micro/nano periodic structures and method for preparing same |
Non-Patent Citations (4)
Title |
---|
Enhanced Fluorescence Microscopic Imaging by Plasmonic Nanostructures: From a 1D Grating to a 2D Nanohole Array;Xiaoqiang Cui等;《Advanced Functional Materials》;20100228;第20卷(第6期);第945-950页 * |
Substrate-supported phospholipid membranes studied by surface plasmon resonance and surface plasmon fluorescence spectroscopy;Tawa, K等;《BIOPHYSICAL JOURNAL》;20051031;第89卷(第4期);第2751页 * |
Tailored Plasmonic Gratings for Enhanced Fluorescence Detection and Microscopic Imaging;Xiaoqiang Cui等;《Advanced Functional Materials》;20100131;第20卷(第4期);第547-553页 * |
ZnO nanorods-enhanced fluorescence for sensitive microarray detection of cancers in serum without additional reporter-amplification;Weihua Hu等;《Biosensors and Bioelectronics》;20110228;第26卷(第8期);第3684页 * |
Also Published As
Publication number | Publication date |
---|---|
CN104880447A (en) | 2015-09-02 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Xia et al. | Use of electroless silver as the substrate in microcontact printing of alkanethiols and its application in microfabrication | |
Notsu et al. | Super-hydrophobic/super-hydrophilic patterning of gold surfaces by photocatalytic lithography | |
CN103991837B (en) | A kind of manufacture method of micro-nano ordered through hole array metal thin film sensor based on piezoelectric substrate thin slice | |
Kang et al. | Protein capture in silica nanotube membrane 3-D microwell arrays | |
CN102901715A (en) | Fluorescence enhanced microarray biochip based on micro/nano periodic structures and method for preparing same | |
CN101617064A (en) | Carry out the universal method of selective area growth of organic molecules by vapour deposition | |
KR101029154B1 (en) | Zinc Oxide Nanostructured Micropattern and Method for Preparing the Same | |
KR100953612B1 (en) | Substrate for immobilizing physiological material, and a method of preparing the same | |
CN104880447B (en) | A kind of ZnO NRs/Au films complex three-dimensional micro-array biochip substrate and preparation method thereof | |
US7491423B1 (en) | Directed spatial organization of zinc oxide nanostructures | |
CN109112601B (en) | Based on TiO2Preparation method and application of Ag nano array photoinduction enhanced Raman substrate | |
KR20070010321A (en) | Method of funtionalized anodic aluminum oxide and method of optical interference bio-sensor using the same and the bio-sensor | |
CN110711610A (en) | Preparation method of super-hydrophobic/super-hydrophilic mixed pattern microarray dendritic gold chip | |
CN105347345A (en) | Silicon micro-nano structure preparing method | |
CN101941672A (en) | Photocatalysis technolog based method for preparing semiconductor nano and metal nano microelectrode array | |
CN101693514A (en) | Method for preparing magnetic suspended coding micro-block array chips and method for applying the preparation method | |
CN102964617A (en) | Film used for fixing biological molecules, and preparation method and applications of film | |
WO2005093416A1 (en) | Substrate for disposing beads and bead disposing method using the same | |
CN106929838A (en) | The method that preparation is suitable to the enhancing substrate of cell surface bio-orthogonal Raman image | |
CN1598575A (en) | Method for making bio-chip | |
CN101145425A (en) | Fe3O4 Au magnetic deposition film preparation method | |
CN1317424C (en) | Preparation process for coating gold by the aid of microwave | |
KR100965238B1 (en) | Substrate for immobilizing physiological material, and a method of preparing the same | |
Li et al. | Fabrication of Honeycomb Gold Arrays for Enhancement of lectrochemical Performance | |
Kamal et al. | Approaches for Sensor Surfaces Modification |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
EXSB | Decision made by sipo to initiate substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant | ||
CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20180213 Termination date: 20190609 |
|
CF01 | Termination of patent right due to non-payment of annual fee |