CN108414492A - It is the method that substrate carries out SERS quantitative analyses using self assembled three-dimensional nanostructure - Google Patents
It is the method that substrate carries out SERS quantitative analyses using self assembled three-dimensional nanostructure Download PDFInfo
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Abstract
It is method that substrate carries out SERS quantitative analyses that the invention discloses a kind of with self assembled three-dimensional nanostructure, steps of the method are：The assembling mode that noble metal nano particles are assisted by interface forms nano particle single layer film on gas-liquid interface；Single layer substrates will be obtained with lifting mode in nano particle single layer film transfer to substrate；After single layer substrates drying, continue to carry out self assembly on substrate with lifting mode, repeats this step until obtaining the substrate of the required nano-particle number of plies；Immersion absorption is carried out to sample to be tested with obtained substrate, substrate is taken out after a certain period of time and is dried, SERS is finally carried out and quantitatively detects.The present invention method is simple, quick, at low cost, high sensitivity, result are accurate and reliable.
Description
Technical field
The invention belongs to nano material and detection technique field, more particularly to a kind of self assembled three-dimensional nanostructure is substrate
The method for carrying out SERS quantitative analyses.
Background technology
Surface enhanced Raman spectroscopy (Surfaced-enhanced Raman Spectroscopy), abbreviation SERS is changing
It learns and materials science field has been developed as a kind of most potential analysis tool.Achieve the effect that enhancing, SERS
The preparation of the preparation method of substrate and effective substrate is as the hot spot studied.Metal nano material is generated with its surface plasma
Electromagnetism amplification, provide effective matrix for SERS, and noble metal nanometer material (being mainly gold, silver, copper) is because of its good light
Property is learned, critical positions are occupied in the field of SERS substrates.
Currently, the preparation method of SERS substrates has sedimentation, template and Langmuir-Blodgett (LB) embrane method.Phase
For first two method, LB embrane methods are more simple and easy to do, and self assembly obtain SERS enhancing substrate in, enhancing effect it is equal
Even property is best.By these assemble methods, base material can be prepared into one-dimensional, two and three dimensions structures.With a Wiki
Bottom is compared, and the solid phase substrate of two and three dimensions overcomes asking for low stability, low homogeneity, easy reunion and hotspot's distribution unevenness
Topic, therefore in practical applications more extensively.In contrast to this, three-dimensional substrates can provide the adsorption area of bigger, highdensity hot spot
(hot spot), therefore produce good SERS enhancing effects.In recent years, three-dimensional assembling SERS substrates have obtained the wide of people
General concern (Tan, R.Z., et al., 3D arrays of SERS substrate for ultrasensitive
molecular detection.Sensors and Actuators A:Physical,2007.139(1-2):p.36-41；
Qian,Y.,et al.,Flexible membranes of Ag-nanosheet-grafted polyamide-
nanofibers as effective 3D SERS substrates.Nanoscale,2014.6(9):p.4781-8.)。
The three-dimensional substrates occurred have porous semi-conductor, the metal nanoparticle that different type metal nanoparticle is modified
Anodised aluminium, metal-modified (Chan, S., et al., the Surface-Enhanced Raman such as nanometer stick array particle
Scattering of Small Molecules from Silver-Coated Silicon Nanopores.Advanced
Materials,2003.15(19):p.1595-1598；Todd L.Williamson,X.G.,Andrew Zukoski,
Aditya Sood,Diego J.D1′az,and Paul W.Bohn,Porous GaN as a template to produce
surface-enhanced Raman scattering-active surfaces.J.Phy.Che B,2005.109:
p.20186-20191；Sinha,G.et al.,Recyclable SERS substrates based on Au-coated
ZnO nanorods.ACS Appl Mater Interfaces,2011.3(7):P.2557-63.), but these three-dimensional substrates still
So there are some disadvantages, such as the substrate of semiconductor, the propagation of light is limited, therefore reduces the increase of their SERS；And
When synthesizing nanometer stick array, multiaperture pellumina is frangible, and extremely hazardous hydrofluoric acid is used in operating process, and oxygen
It is all unstable in strong acid and highly basic to change zinc array.
In addition, the qualitative analysis that SERS is used for substance earliest has begun persistently to step on the road of quantitative analysis in recent years
Into the quantitative analysis tech of SERS spectra also begins to be widely used in the necks such as environmental analysis, food security, biological medicine analysis
Domain.
To sum up, although the method for preparation substrate and quantitative analysis achieve some achievements at present, it is used as receiving for substrate
The enhancement effect of rice material different loci is also different, therefore, the overriding challenges of SERS quantitative analyses be exactly substrate uniformity and
Reliability.
Invention content
It is that substrate carries out SERS quantitative analyses that the purpose of the present invention is to provide a kind of using self assembled three-dimensional nanostructure
Method.The present invention method is simple, quick, at low cost, high sensitivity, result are accurate and reliable, can be used for quantitative analysis.
To achieve the above object, the present invention takes following technical scheme：
A method of SERS quantitative analyses being carried out for substrate using self assembled three-dimensional nanostructure, are included the following steps：
(1) noble metal nano particles are formed into nano particle single layer by the assembling mode that interface assists on gas-liquid interface
Film；
(2) single layer substrates will be obtained in nano particle single layer film transfer to substrate with lifting mode；
(3) after single layer substrates drying, continue to carry out self assembly on substrate with lifting mode, repeat this step until obtaining
To the substrate of the required nano-particle number of plies；
(4) substrate obtained with step (3) carries out immersion absorption to sample to be tested, after certain time (general 2~3 hours)
It takes out substrate to be dried, finally carries out SERS and quantitatively detect.
The nano-particle of interfacial assembly substrate is variously-shaped, size noble metal nano particles, it is preferred that described expensive
Metal nanoparticle includes the sheet of gold or silver, rodlike, spherical, rodlike, starlike, cubic or rectangular-shape nano-particle.
In the above-mentioned methods, the specific method is as follows for step (1) the preparation nano particle single layer film：Nano-particle is molten
Drop is added in the container containing liquid phase, and nano particle single layer is formed in gas-liquid interface over-assemble in such a way that interface assists
Film.
Preferably, the container is beaker, vial, centrifuge tube lid or culture dish.
Preferably, the liquid phase is polar solvent；It is furthermore preferred that the liquid phase is water, ethylene glycol or diethylene glycol.
Preferably, the nano-particle solution is the non-polar solution of nano-particle, and the non-polar solution is selected from dichloro
It is one or more in methane, hexamethylene, octane.
Preferably, the volume of the nano-particle solution and the proportional increase of the diameter of container.It is furthermore preferred that when container is straight
When diameter is 2cm, nano-particle solution volume is 200 μ l.
In the above-mentioned methods, the substrate of the step (2) is preferably silicon chip or glass slide.
In the above-mentioned methods, sample to be tested is biomolecule, pesticide, illegal drug or food additives.
The present invention is finally arranged single layer and multilayer by way of a kind of LBL self-assembly of simple and quick liquid level auxiliary
The nano-particle of row is transferred on clean silicon chip and/or glass slide, and obtained substrate is relatively uniform, so in binding molecule
When can be effectively accomplished uniform absorption so that it is essentially identical to put binding molecule number each of in substrate, in table
Reach the requirement of quantitative detection in face enhancing Raman detection.
The present invention has the following advantages that compared with prior art：
(1) method that the present invention prepares self assembled three-dimensional nanostructure substrate is prepared relative to template and sedimentation
For 3D substrates, operation is easier, and the uniformity is higher.
(2) method that the present invention prepares self assembled three-dimensional nanostructure substrate is applicable not only to Silver nanorod and golden octahedral
Body is also applied for other variously-shaped noble metal nano particles, widely applicable.
(3) the self assembled three-dimensional nanostructure substrate prepared by the present invention, sensitivity higher.
(4) the self assembled three-dimensional nanostructure substrate prepared by the present invention is applicable not only to a certain in terms of Molecular Detection
The quantitative detection of the single molecule of kind, moreover it is possible to be suitable for the quantitative detection of other and human health environmental correclation molecule.
Description of the drawings
Fig. 1 is the preparation flow schematic diagram of 3-D nano, structure substrate of the present invention；
The 6 layers of Silver nanorod substrate surface SEM (A) and section that Fig. 2, which is the liquid phase of interfacial assembly of the present invention, to be assembled when being water
SEM(B)；
1 layer of substrate (A) that Fig. 3, which is the liquid phase of interfacial assembly of the present invention, to be assembled when being ethylene glycol, 2 layers (B), 3 layers (C), 4 layers
(D), 5 layers (E), 6 layers (F), 7 layers (G), 8 layers (H), 9 layers (I), 10 layers (J), 12 layers (K), 14 layers (L), 16 layers (M), 18 layers
(N), the surface of 20 layers (O) and section SEM；
The octahedral surface of 1-10 layer gold and section that Fig. 4, which is the liquid phase of interfacial assembly of the present invention, to be assembled when being diethylene glycol
SEM；
Fig. 5 is the spectrogram (A) and linear relationship that the three-dimensional Silver nanorod substrate that the present invention assembles quantitatively detects urea molecule
Scheme (B)；
Fig. 6 is the spectrogram (A) and linearly that the three Vygen octahedron substrates that assemble of the present invention quantitatively detect phenylalanine molecule
Relational graph (B).
Specific implementation mode
Following specific examples is the further explanation to method provided by the invention and technical solution, but is not construed as
Limitation of the present invention.
Embodiment 1：
The liquid phase of the preparation of Silver nanorod three-dimensional substrates, interfacial assembly is water.
Fig. 1 provides the preparation flow schematic diagram of three-dimensional substrates.
Specifically preparation method is：
200 μ l Silver nanorods (dichloromethane solution) are taken uniformly to be mixed with 200 μ l dichloromethane, 200 μ l hexamethylenes, then
30 μ l octanes are added uniformly to mix, 200 μ l of its mixed liquor is taken to be added dropwise on the water surface of diameter 2cm, after organic phase is fully volatilized,
Clean silicon chip and/or glass slide is sunken under liquid level, and the single layer Silver nanorod on the water surface is made to be transferred to by way of lifting
On silicon chip or glass slide, the single layer substrates of Silver nanorod are obtained, at room temperature after the liquid drying on silicon chip or glass slide, are continued
It assembles on it in the same manner, obtains the substrate of the Silver nanorod of 6 layers of assembling.The SEM characterizations of 6 layers of assembling substrate are shown in Fig. 2.
Embodiment 2：
The liquid phase of the preparation of Silver nanorod single layer and multi-layer three-dimension substrate, interfacial assembly is ethylene glycol
Fig. 1 provides the preparation flow schematic diagram of Silver nanorod single layer and multi-layer substrate.
Specifically preparation method is：Take 200 μ l Silver nanorods (dichloromethane solution) that the ethylene glycol liquid level in diameter 2cm is added dropwise
On, after organic phase is fully volatilized, clean silicon chip and/or glass slide are sunken under liquid level, and liquid level is made by way of lifting
On single layer Silver nanorod be transferred on silicon chip or glass slide, obtain the single layer substrates of Silver nanorod, substrate, which is placed in vacuum, to be done
Dry 30min continues after the liquid drying on silicon chip or glass slide with phase Tongfang in dry case and/or lyophilizer
Formula assembles on it, obtains the substrate of the Silver nanorod of 1 layer to 20 layers assembling.The SEM characterizations of 1-20 layers of assembling substrate are shown in Fig. 3.
Embodiment 3：
The liquid phase of the preparation of golden octahedron single layer and multi-layer three-dimension substrate, interfacial assembly is diethylene glycol
Fig. 1 provides single layer and the preparation flow schematic diagram of the octahedra substrate of multilayer gold.Specifically preparation method is：
It takes 200 μ l gold octahedra (dichloromethane solution) to be added dropwise on the diethylene glycol liquid level of diameter 2cm, waits for that organic phase is filled
After dividing volatilization, clean silicon chip and/or glass slide are sunken under liquid level, and the single layer of gold octahedral on liquid level is made by way of lifting
Body is transferred on silicon chip or glass slide, obtains the octahedral single layer substrates of gold, and substrate is placed in vacuum drying chamber and/or freezing is true
Dry 30min in empty drier, after the liquid drying on silicon chip or glass slide, continuation assembles on it in the same manner, obtains
The octahedral substrate of gold assembled to 1 layer to 10 layers.The SEM characterizations of 1-10 layers of assembling substrate are shown in Fig. 4.
Embodiment 4：
The SERS quantitative analyses of Silver nanorod three-dimensional substrates：
(1) by taking Silver nanorod three-dimensional substrates as an example, urea molecule carries out SERS quantitative analysis detections as analyte.
(2) 200 μ l Silver nanorods (dichloromethane solution) is taken uniformly to be mixed with 200 μ l dichloromethane and 200 μ l hexamethylenes,
Then 30 μ l octanes are added uniformly to mix, 200 μ l of this mixed liquor are added dropwise on the water surface of a diameter of 2cm, wait for that organic phase is abundant
After volatilization, silicon chip is sunken under liquid level, is transferred to the Silver nanorod on liquid level thereon by the way of lifting, after drying at room temperature
Continuation assembles by this method, until being assembled into 6 layers.
(3) by prepare 6 layers of substrate be separately immersed in various concentration urea liquid (300mM, 100mM, 50mM,
30mM, 5mM, 1mM, 250 μM, 125 μM) in, take out drying after 2 hours, carrying out SERS detections specifically will be adsorbed with difference
The substrate of the urea molecule of concentration is respectively placed in progress SERS imagings under confocal microscope, and 532nm exciting lights excite work(
Rate is 3.07mW, time of integration 10s, and the object lens for being 0.25 by 10 times of NA values, which focus on SERS substrate over-assembles, silver nanoparticle
The one side of stick, the Raman photon scattered are collected by the object lens, after optical filter and grating beam splitting, on CCD
To the Surface enhanced Raman spectroscopy of SERS substrate urea molecules.Fig. 5 A are the SERS spectra figure of various concentration urea molecule, Fig. 5 B
It is that (absorption peak is in 1004cm for various concentration urea molecule-1) with the relational graph of SERS intensity, as seen from the figure, R2=0.980, have very
Good linear relationship, there is certain feasibility and reliability for the quantitative detection of molecule.
Embodiment 5：The SERS quantitative analyses of golden octahedron three-dimensional substrates
(1) by taking the octahedra three-dimensional substrates of above-mentioned gold as an example, phenylalanine molecule carries out SERS quantitative analyses as analyte
Detection.
(2) 200 μ l gold octahedra (nonpolar solvent) are taken uniformly to be mixed with 200 μ l dichloromethane and 200 μ l hexamethylenes, so
After 30 μ l octanes be added uniformly mix, 200 μ l of this mixed liquor are added dropwise on the water surface of a diameter of 2cm, wait for that organic phase is fully waved
After hair, silicon chip is sunken under liquid level, is transferred to the golden octahedron on liquid level thereon by the way of lifting, drying at room temperature is subsequent
It is continuous to assemble by this method, until being assembled into 5 layers.
(3) prepare 5 layers of substrate are separately immersed in phenylalanine (1mM, 500 μM, 200 μM, 100 μ of various concentration
M, 50 μM, 30 μM, 20 μM, 15 μM) in, take out drying within 2 hours, carrying out SERS detections specifically will be adsorbed with various concentration
The substrate of phenylalanine molecule is respectively placed in progress SERS imagings, 785nm exciting lights, exciting power under confocal microscope
For 4.34mW, time of integration 10s, the object lens for being 0.25 by 10 times of NA values, which focus on SERS substrate over-assembles, golden octahedron
One side, the Raman photon scattered is collected by the object lens, after optical filter and grating beam splitting, obtained on CCD
The Surface enhanced Raman spectroscopy of SERS substrate phenylalanine molecules.Fig. 6 A are the SERS spectras of various concentration phenylalanine molecule
Figure, Fig. 6 B are that (absorption peak is in 1000cm for various concentration phenylalanine molecule-1) with the relational graph of SERS intensity, as seen from the figure, R2
=0.990, there is good linear relationship, there is certain feasibility and reliability for the quantitative detection of molecule.
The explanation of above example is only intended to help to understand the method for the present invention and its core concept.It should be pointed out that for
For those skilled in the art, without departing from the principle of the present invention, if can also be carried out to the present invention
Dry improvement and modification, these improvement and modification are also fallen into the claimed scope of the invention.
Claims (10)
1. a kind of utilizing the method that self assembled three-dimensional nanostructure is substrate progress SERS quantitative analyses, which is characterized in that including
Following steps：
(1) assembling mode for assisting noble metal nano particles by interface forms nano particle single layer film on gas-liquid interface；
(2) single layer substrates will be obtained in nano particle single layer film transfer to substrate with lifting mode；
(3) after single layer substrates drying, continue to carry out self assembly on substrate with lifting mode, repeat this step until obtaining institute
Need the substrate of the nano-particle number of plies；
(4) substrate obtained with step (3) carries out immersion absorption to sample to be tested, takes out substrate after a certain period of time and is dried,
SERS is finally carried out quantitatively to detect.
2. according to the method described in claim 1, it is characterized in that, the noble metal include gold or silver, the nano-particle
Shape is selected from sheet, rodlike, spherical, rodlike, starlike, cubic or rectangular-shape.
3. according to the method described in claim 1, it is characterized in that, the step (1) prepares the specific of nano particle single layer film
Method is as follows：Nano-particle solution is added drop-wise in the container containing liquid phase, in such a way that interface assists on gas-liquid interface
Assembling forms nano particle single layer film.
4. according to the method described in claim 3, it is characterized in that, the container is beaker, vial, centrifuge tube lid or training
Support ware.
5. according to the method described in claim 3, it is characterized in that, the liquid phase is polar solvent.
6. according to the method described in claim 5, it is characterized in that, the polar solvent is water, ethylene glycol or diethylene glycol.
7. according to the method described in claim 3, it is characterized in that, the nano-particle solution is the nonpolar molten of nano-particle
Liquid, the non-polar solution are one or more in dichloromethane, hexamethylene, octane.
8. according to the method described in claim 3, it is characterized in that, the nano-particle solution volume and container diameter at
Ratio increases.
9. according to the method described in claim 1, it is characterized in that, the substrate of the step (2) is silicon chip or glass slide.
10. according to the method described in claim 1, it is characterized in that, the sample to be tested is biomolecule, pesticide, illicit drug
Product or food additives.
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Cited By (7)
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| CN109239051A (en) * | 2018-09-29 | 2019-01-18 | 东南大学 | Transferable type surface-enhanced Raman detection substrate of a kind of flexibility and its preparation method and application |
| CN109612976A (en) * | 2018-12-24 | 2019-04-12 | 北华大学 | A kind of 3-dimensional multi-layered body structure surface enhancing Raman substrate and preparation method thereof |
| CN110790220A (en) * | 2019-10-29 | 2020-02-14 | 深圳大学 | Surface-enhanced Raman scattering substrate, preparation method thereof and in-situ rapid detection method |
| CN110823859A (en) * | 2019-11-05 | 2020-02-21 | 临沂大学 | High-sensitivity Raman signal detection substrate for urea and preparation method and application thereof |
| CN111017872A (en) * | 2019-11-25 | 2020-04-17 | 西安邮电大学 | Preparation method of sandwich structure composite nano array substrate |
| CN112893865A (en) * | 2021-03-26 | 2021-06-04 | 江苏师范大学 | Double-layer gold nanoparticle modified flexible SERS substrate and preparation method thereof |
| WO2021179347A1 (en) * | 2020-03-10 | 2021-09-16 | 中国科学院城市环境研究所 | Raman spectrum-based method for measuring free chlorine in water |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109239051A (en) * | 2018-09-29 | 2019-01-18 | 东南大学 | Transferable type surface-enhanced Raman detection substrate of a kind of flexibility and its preparation method and application |
| CN109612976A (en) * | 2018-12-24 | 2019-04-12 | 北华大学 | A kind of 3-dimensional multi-layered body structure surface enhancing Raman substrate and preparation method thereof |
| CN110790220A (en) * | 2019-10-29 | 2020-02-14 | 深圳大学 | Surface-enhanced Raman scattering substrate, preparation method thereof and in-situ rapid detection method |
| CN110823859A (en) * | 2019-11-05 | 2020-02-21 | 临沂大学 | High-sensitivity Raman signal detection substrate for urea and preparation method and application thereof |
| CN111017872A (en) * | 2019-11-25 | 2020-04-17 | 西安邮电大学 | Preparation method of sandwich structure composite nano array substrate |
| CN111017872B (en) * | 2019-11-25 | 2022-03-04 | 西安邮电大学 | Preparation method of sandwich structure composite nano array substrate |
| WO2021179347A1 (en) * | 2020-03-10 | 2021-09-16 | 中国科学院城市环境研究所 | Raman spectrum-based method for measuring free chlorine in water |
| CN112893865A (en) * | 2021-03-26 | 2021-06-04 | 江苏师范大学 | Double-layer gold nanoparticle modified flexible SERS substrate and preparation method thereof |
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Application publication date: 20180817 |