CN108226133A - It is a kind of for local Electromagnetic enhancement device of Raman Characterization and preparation method thereof, application and application method - Google Patents
It is a kind of for local Electromagnetic enhancement device of Raman Characterization and preparation method thereof, application and application method Download PDFInfo
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Abstract
The present invention relates to a kind of for local Electromagnetic enhancement device of Raman Characterization and preparation method thereof, application and application method, belong to detection technique field, which includes composition metal nanohole array substrate and atomic force microscope probe;Composition metal nanohole array substrate is made of from bottom to top the silver layer of hard substrate, layers of chrome, layer gold, upper surface cladding golden film successively, nanohole array is set on silver layer and layer gold, and nano-pore vertical depth is less than the sum of silver layer and the thickness of layer gold, nano-pore hole wall is coated with golden film;Atomic force microscope probe surface is coated with silverskin, and probe tip surface has striped.The device can realize the molecular fingerprint spectral characterization analysis of unimolecule rank, and then normal Raman or surface enhanced is overcome to scatter Raman can not obtain true unimolecule the configuration conformational difference of substrate surface the shortcomings that, the characterization of isomer molecule configuration conformation can also be applied to, realizes the discriminant analysis of the isomers on single molecules level.
Description
Technical field
The invention belongs to detection technique fields, and in particular to a kind of local Electromagnetic enhancement device for Raman Characterization
Part and preparation method thereof, application and application method.
Background technology
Surface Enhanced Raman Scattering Spectrum technology has the enhancement effect of electromagnetic field by substrate material surface, can be greatly
Enhance the Raman scattering signal of trace molecules, the structure of molecule can be obtained by its feature peak position, intensity or line width information difference
The difference of type conformation and its discriminating of isomer have in the quick analysis of trace amount environment pollutant and its different isomer
There is unique advantage, especially played a very important role in the analysis of the isomer molecule with different toxicity.Due to
Surface Enhanced Raman Scattering Spectrum technology is difficult to obtain in spatial resolution and the limitation of local Electromagnetic enhancement aspect of performance
Raman scattering signal on unimolecule scale, so nanometer pinpoint increases with the needle point Raman that nanostructured local enhancing technology is combined
Strong technology has started the phenetic analysis of the material for molecular scale, biotic environment sample, but the technology is still difficult to obtain
Otherness between unimolecule configuration conformation and its isomer.Therefore, urgent need one kind can obtain true unimolecule and serve as a contrast
The device and characterizing method of the configuration conformational difference of bottom surface.
Invention content
In view of this, one of the objects of the present invention is to provide a kind of local Electromagnetic enhancements for Raman Characterization
Device;The second object of the present invention is to provide a kind of preparation side of the local Electromagnetic enhancement device for Raman Characterization
Method;The third object of the present invention is to provide a kind of local Electromagnetic enhancement device for Raman Characterization in unimolecule structure
Application in type conformation;The fourth object of the present invention is to provide a kind of local Electromagnetic enhancement device for Raman Characterization
The application method of part.
In order to achieve the above objectives, the present invention provides following technical solution:
1st, a kind of local Electromagnetic enhancement device for Raman Characterization, including composition metal nanohole array substrate
And atomic force microscope probe;The composition metal nanohole array substrate is from bottom to top successively by hard substrate, layers of chrome, gold
The silver layer composition of layer, upper surface cladding golden film, the nanohole array is set on silver layer and layer gold, and nano-pore vertical depth
Less than the sum of silver layer and the thickness of layer gold, nano-pore hole wall is coated with golden film;The atomic force microscope probe surface is coated with
Silverskin, probe tip surface have striped.
Further, the hard substrate is silicon chip or sheet glass.
Further, the thickness of the layers of chrome is 2-3nm, and the thickness of layer gold is 50-100nm, and the thickness of silver layer is 20-
50nm, the thickness of silverskin is 10-20nm.
Further, nano-pore aperture is 50-100nm, period 100-200nm in the nanohole array.
Further, the material of the silverskin is doping and the irregular ag material of lattice.
Further, radius of curvature is less than 30nm at the probe tip.
2nd, the preparation method of a kind of local Electromagnetic enhancement device for Raman Characterization, the method packet
Include following steps:
(1) composition metal nanohole array substrate is prepared
After hard substrate is cleaned up, it is sequentially depositing layers of chrome, layer gold and silver layer from bottom to top, then utilizes focused ion
Beam lithographic technique etches silver layer first, and the silver atoms sputtered in etching process are redeposited in the place that silver layer is not etched,
After the silver layer is cut through, start to etch layer gold, the gold atom sputtered in etching process is redeposited not to be carved in silver layer
On the place of erosion and the hole wall of etched formed nano-pore, until the place that silver layer is not etched is received with etched formed
The gold atom that the hole wall of metre hole is sputtered out is completely covered and nano-pore vertical depth is less than the sum of silver layer and the thickness of layer gold, system
Obtain composition metal nanohole array substrate;
(2) needle point is prepared
After atomic force microscope probe is cleaned up, silverskin is deposited in detecting probe surface, is then carved using focused ion beam
Erosion technology handles probe tip, and needle surface is made to have striped.
Further, described method includes following steps:
(1) composition metal nanohole array substrate is prepared
After using plasma washing machine or chemical method, hard substrate is cleaned up, steamed using magnetron sputtering or vacuum
Coating technology is sequentially depositing layers of chrome, layer gold and silver layer from bottom to top, then using focused-ion-beam lithography technology, using gallium source, choosing
With 15 μm of diaphragms, it is less than 5pA in line, focused ion beam dose is 0.02-0.1nC/ μm2Under the conditions of, silver layer is etched first, is carved
It is redeposited in the place that silver layer is not etched to lose the silver atoms that sputter in the process, after the silver layer is cut through, starts to carve
Layer gold is lost, the gold atom sputtered in etching process is redeposited to be received in the place that silver layer is not etched with etched formed
On the hole wall of metre hole, until the place that silver layer is not etched and the gold atom that the hole wall of etched formed nano-pore is sputtered out
It is completely covered and nano-pore vertical depth is less than the sum of silver layer and the thickness of layer gold, composition metal nanohole array substrate is made;
(2) needle point is prepared
After using plasma washing machine, atomic force microscope probe is cleaned up, using magnetron sputtering or vacuum evaporation
Silverskin is deposited in detecting probe surface, then using focused-ion-beam lithography technology, using helium source, 15 μm of diaphragms is selected, is in line
1pA, focused ion beam dose are 0.01-0.02nC/ μm2Under the conditions of, probe tip is etched, adjusts the radius of curvature of needle point, and
Striped is etched in needle surface.
3rd, a kind of local Electromagnetic enhancement device for Raman Characterization is characterized in unimolecule Raman fingerprint
In application.
4th, the application method of a kind of local Electromagnetic enhancement device for Raman Characterization, the method tool
Body is:By sample to be tested drop on the composition metal nanohole array substrate, then utilize simultaneously described in being equipped with
The atomic force microscope and Raman common focusing micro spectrum instrument of atomic force microscope probe are to composition metal nanohole array substrate
Surface is scanned, on comparative analysis composition metal nanohole array substrate in nano-pore, between two adjacent nano holes, three
Needle point enhancing scattered signal feature peak position and peak intensity between adjacent nano hole, obtain single testing molecule in substrate surface configuration
The difference of conformation.
The beneficial effects of the present invention are:The present invention provides a kind of local Electromagnetic enhancements for Raman Characterization
Device and preparation method thereof, application and application method, the device include composition metal nanohole array substrate and atomic force microscopy
Mirror probe, the layers of chrome deposited in the composition metal nanohole array substrate can improve adsorptivity of the layer gold to hard substrate, sink
Long-pending silver layer can greatly enhance monomolecular Raman scattering signal near nano-pore, simultaneously because golden film is coated on silver layer table
Face can be effectively prevented silver layer oxidation.It, can be to substrate by controlling nanoporous size, period on nanohole array substrate
Surface local magnetic distribution is adjusted and the capture of individual molecule, in conjunction with the surface local electromagnetism of gold, silver noble metal film
Enhancing effect can greatly enhance in nano-pore, the Raman scattering signal of the molecule of array surface attachment.By atom in the present invention
Force microscope detecting probe surface coats silverskin, and radius of curvature at probe tip is made to be less than 30nm so that needle point local electromagnetic field increases
It is potent to be confined to unimolecule size, while striped is etched around needle point, due to the material doped of detecting probe surface silverskin and
Lattice is irregular, can influence helium ion etching speed, causes the fringe depths etched inconsistent, can greatly increase needle in this way
Sharp strong local electromagnetic field.The device can realize the phenetic analysis of the molecular fingerprint spectrum of unimolecule rank, and then overcome common
Raman or surface enhanced scattering Raman can not obtain true unimolecule the configuration conformational difference of substrate surface the shortcomings that, together
When, the characterization of isomer molecule configuration conformation can also be applied to, realizes the discriminant analysis of the isomers on single molecules level.
Description of the drawings
In order to make the purpose of the present invention, technical solution and advantageous effect clearer, the present invention provides drawings described below and carries out
Explanation:
Fig. 1 is the process flow chart for preparing composition metal nanohole array substrate;
Fig. 2 is focused-ion-beam lithography schematic diagram when preparing composition metal nanohole array substrate;
Fig. 3 is the vertical view of composition metal nanohole array substrate;
Fig. 4 is atomic force microscope probe needle point schematic diagram;
Fig. 5 is when utilizing atomic force microscope probe scanning composition metal nano-pore array structure different location in the present invention
Schematic diagram;
Wherein, in Fig. 1,1 represents hard substrate, and 2 represent layers of chrome, and 3 represent layer gold, and 4 represent silver layer, and 5 represent golden film, 6 tables
Show nano-pore.
Specific embodiment
The preferred embodiment of the present invention will be described in detail below.
Embodiment 1
Prepare the local Electromagnetic enhancement device for Raman Characterization
(1) composition metal nanohole array substrate is prepared
Referring to composition metal nanohole array underlay producing technique flow chart in Fig. 1, using plasma washing machine by glass
After glass piece 1 cleans up, it is sequentially depositing the layers of chrome 2 of 2nm thickness, 3 and of layer gold of 100nm thickness from bottom to top using vacuum evaporation technology
The silver layer 4 of 20nm thickness then using focused-ion-beam lithography technology (etching principle is referring to Fig. 2), using gallium source, selects 15 μm of light
Door screen is less than 5pA in line, and focused ion beam dose is 0.1nC/ μm2Under the conditions of, silver layer 4 is etched first, is sputtered in etching process
The silver atoms that go out are redeposited in the place that silver layer 4 is not etched, after the silver layer 4 is cut through, start to etch layer gold 3, carve
It is redeposited in the place that silver layer 4 is not etched and etched formed nano-pore 6 to lose the gold atom sputtered in the process
On hole wall, until the place that silver layer 4 is not etched and the gold atom that the hole wall of etched formed nano-pore 6 is sputtered out are complete
It covers and 6 vertical depth of nano-pore is less than the sum of silver layer 4 and the thickness of layer gold 3, form golden film 5 on silver layer 4 at this time, be made and receive
Metre hole aperture is 50nm, and the period is the composition metal nanohole array substrate of 100nm, the composition metal nanohole array substrate
Vertical view is as shown in Figure 3;
(2) needle point is prepared
After using plasma washing machine, atomic force microscope probe is cleaned up, with doping and the irregular silver of lattice
Material is coating materials, using vacuum evaporation detecting probe surface deposition 20nm thickness silverskin, then using focused-ion-beam lithography technology,
Using helium source, 15 μm of diaphragms are selected, are 1pA in line, focused ion beam dose is 0.02nC/ μm2Under the conditions of, etch probe needle
Point, it is 20nm, and etch striped in needle surface to make radius of curvature at needle point, and prepared atomic force microscope probe needle point shows
It is intended to as shown in Figure 4.
Embodiment 2
Prepare the local Electromagnetic enhancement device for Raman Characterization
(1) composition metal nanohole array substrate is prepared
Referring to composition metal nanohole array underlay producing technique flow chart in Fig. 1, using plasma washing machine by glass
After glass piece 1 cleans up, it is sequentially depositing the layers of chrome 2 of 3nm thickness, 3 and of layer gold of 50nm thickness from bottom to top using vacuum evaporation technology
The silver layer 4 of 35nm thickness then using focused-ion-beam lithography technology (etching principle is referring to Fig. 2), using gallium source, selects 15 μm of light
Door screen is less than 5pA in line, and focused ion beam dose is 0.05nC/ μm2Under the conditions of, silver layer 4 is etched first, is splashed in etching process
The silver atoms of injection are redeposited in the place that silver layer 4 is not etched, after the silver layer 4 is cut through, start to etch layer gold 3,
The gold atom sputtered in etching process is redeposited in the place that silver layer 4 is not etched and etched formed nano-pore 6
Hole wall on, until place and the gold atom that is sputtered out of hole wall of etched formed nano-pore 6 that silver layer 4 is not etched are complete
All standing and 6 vertical depth of nano-pore are less than the sum of silver layer 4 and the thickness of layer gold 3, form golden film 5 on silver layer 4 at this time, are made
Nano-pore aperture be 80nm, the period be 150nm composition metal nanohole array substrate, the composition metal nanohole array substrate
Vertical view it is as shown in Figure 3;
(2) needle point is prepared
After using plasma washing machine, atomic force microscope probe is cleaned up, with doping and the irregular silver of lattice
Material is coating materials, using vacuum evaporation detecting probe surface deposition 15nm thickness silverskin, then using focused-ion-beam lithography technology,
Using helium source, 15 μm of diaphragms are selected, are 1pA in line, focused ion beam dose is 0.01nC/ μm2Under the conditions of, etch probe needle
Point, it is 25nm, and etch striped in needle surface to make radius of curvature at needle point, and prepared atomic force microscope probe needle point shows
It is intended to as shown in Figure 4.
Embodiment 3
Prepare the local Electromagnetic enhancement device for Raman Characterization
(1) composition metal nanohole array substrate is prepared
Referring to composition metal nanohole array underlay producing technique flow chart in Fig. 1, using plasma washing machine by glass
After glass piece 1 cleans up, it is sequentially depositing the layers of chrome 2 of 2nm thickness, 3 and of layer gold of 75nm thickness from bottom to top using vacuum evaporation technology
The silver layer 4 of 50nm thickness then using focused-ion-beam lithography technology (etching principle is referring to Fig. 2), using gallium source, selects 15 μm of light
Door screen is less than 5pA in line, and focused ion beam dose is 0.02nC/ μm2Under the conditions of, silver layer 4 is etched first, is splashed in etching process
The silver atoms of injection are redeposited in the place that silver layer 4 is not etched, after the silver layer 4 is cut through, start to etch layer gold 3,
The gold atom sputtered in etching process is redeposited in the place that silver layer 4 is not etched and etched formed nano-pore 6
Hole wall on, until place and the gold atom that is sputtered out of hole wall of etched formed nano-pore 6 that silver layer 4 is not etched are complete
All standing and 6 vertical depth of nano-pore are less than the sum of silver layer 4 and the thickness of layer gold 3, form golden film 5 on silver layer 4 at this time, are made
Nano-pore aperture is 100nm, and the period is the composition metal nanohole array substrate of 200nm, which serves as a contrast
The vertical view at bottom is as shown in Figure 3;
(2) needle point is prepared
After using plasma washing machine, atomic force microscope probe is cleaned up, with doping and the irregular silver of lattice
Material is coating materials, using vacuum evaporation detecting probe surface deposition 10nm thickness silverskin, then using focused-ion-beam lithography technology,
Using helium source, 15 μm of diaphragms are selected, are 1pA in line, focused ion beam dose is 0.02nC/ μm2Under the conditions of, etch probe needle
Point, it is 27nm, and etch striped in needle surface to make radius of curvature at needle point, and prepared atomic force microscope probe needle point shows
It is intended to as shown in Figure 4.
Embodiment 4
Single Microcystin molecule is characterized in the difference of substrate surface configuration conformation
The device prepared in embodiment 1 is used for difference table of the single Microcystin molecule in substrate surface configuration conformation
Sign, it is specific as follows:
Compound concentration is 10-12The Microcystin the sample solution to be tested of M takes 10 μ L sample to be tested drops to be prepared in embodiment 1
Composition metal nanohole array substrate on, then using being equipped in embodiment 1 that treated, atomic force microscope is visited simultaneously
The atomic force microscope and Raman common focusing micro spectrum instrument of needle are scanned composition metal nanohole array substrate surface, borrow
Position of the atomic force microscope control probe tip on composition metal nano-pore array structure is helped, as shown in figure 5, and then obtaining
The pinpoint enhanced Raman scattered signal of Microcystin molecule, comparative analysis are compound on composition metal nano-pore array structure substrate
Needle point enhancing in metal nano-void array substrate in nano-pore, between two adjacent nano holes, between three adjacent nano holes
Scattered signal feature peak position and peak intensity, obtain the lying low of single Microcystin molecule, molecule insertion nano-pore, molecular atoms with
The difference of configuration conformation between gold bonding and Microcystin molecule isomer molecule.
Embodiment 5
Isomers of carotenoids is characterized in the difference of substrate surface configuration conformation
The device prepared in embodiment 1 is characterized for difference of the isomers of carotenoids in substrate surface configuration conformation,
It is specific as follows:
Compound concentration is 10-12The isomers of carotenoids the sample solution to be tested of M takes 10 μ L sample to be tested drops in embodiment 1
On the composition metal nanohole array substrate of middle preparation, then using being equipped in embodiment 1 that treated, atomic force is shown simultaneously
The atomic force microscope and Raman common focusing micro spectrum instrument of micro mirror probe carry out composition metal nanohole array substrate surface
Scanning, by position of the atomic force microscope control probe tip on composition metal nano-pore array structure, as shown in figure 5,
And then obtain the pinpoint enhanced Raman scattering letter of isomers of carotenoids molecule on composition metal nano-pore array structure substrate
Number, on comparative analysis composition metal nanohole array substrate in nano-pore, between two adjacent nano holes, three adjacent nano holes
Between needle point enhancing scattered signal feature peak position and peak intensity, obtain the difference of configuration conformation between isomers of carotenoids molecule
It is different.
Finally illustrate, preferred embodiment above is merely illustrative of the technical solution of the present invention and unrestricted, although logical
It crosses above preferred embodiment the present invention is described in detail, however, those skilled in the art should understand that, can be
Various changes are made to it in form and in details, without departing from claims of the present invention limited range.
Claims (10)
1. a kind of local Electromagnetic enhancement device for Raman Characterization, which is characterized in that including composition metal nano-pore
Array substrate and atomic force microscope probe;The composition metal nanohole array substrate from bottom to top successively by hard substrate,
Layers of chrome, layer gold, upper surface cladding golden film silver layer composition, the nanohole array is set on silver layer and layer gold, and nano-pore
Vertical depth is less than the sum of silver layer and the thickness of layer gold, and nano-pore hole wall is coated with golden film;The atomic force microscope probe table
Bread is covered with silverskin, and probe tip surface has striped.
2. a kind of local Electromagnetic enhancement device for Raman Characterization according to claim 1, which is characterized in that
The hard substrate is silicon chip or sheet glass.
3. a kind of local Electromagnetic enhancement device for Raman Characterization according to claim 1, which is characterized in that
The thickness of the layers of chrome is 2-3nm, and the thickness of layer gold is 50-100nm, and the thickness of silver layer is 20-50nm, and the thickness of silverskin is
10-20nm。
4. a kind of local Electromagnetic enhancement device for Raman Characterization according to claim 1, which is characterized in that
Nano-pore aperture is 50-100nm, period 100-200nm in the nanohole array.
5. a kind of local Electromagnetic enhancement device for Raman Characterization according to claim 1, which is characterized in that
The material of the silverskin is doping and the irregular ag material of lattice.
6. a kind of local Electromagnetic enhancement device for Raman Characterization according to claim 1, which is characterized in that
Radius of curvature is less than 30nm at the probe tip.
7. a kind of preparation side of local Electromagnetic enhancement device for Raman Characterization of claim 1-6 any one of them
Method, which is characterized in that described method includes following steps:
(1) composition metal nanohole array substrate is prepared
After hard substrate is cleaned up, it is sequentially depositing layers of chrome, layer gold and silver layer from bottom to top, is then carved using focused ion beam
Erosion technology etches silver layer first, and the silver atoms sputtered in etching process are redeposited in the place that silver layer is not etched, treat institute
It states after silver layer cut through, starts to etch layer gold, the gold atom sputtered in etching process is redeposited not to be etched in silver layer
On the hole wall of local and etched formed nano-pore, until local and etched the formed nano-pore that silver layer is not etched
The gold atom that is sputtered out of hole wall be completely covered and nano-pore vertical depth is less than the sum of silver layer and the thickness of layer gold, be made multiple
Alloy category nanohole array substrate;
(2) needle point is prepared
After atomic force microscope probe is cleaned up, silverskin is deposited in detecting probe surface, then utilizes focused-ion-beam lithography skill
Art handles probe tip, and needle surface is made to have striped.
8. preparation method according to claim 7, which is characterized in that described method includes following steps:
(1) composition metal nanohole array substrate is prepared
After using plasma washing machine or chemical method, hard substrate is cleaned up, using magnetron sputtering or vacuum evaporation skill
Art is sequentially depositing layers of chrome, layer gold and silver layer from bottom to top, then using focused-ion-beam lithography technology, using gallium source, selects 15 μ
M diaphragms are less than 5pA in line, and focused ion beam dose is 0.02-0.1nC/ μm2Under the conditions of, silver layer is etched first, it is etched
The silver atoms that are sputtered in journey are redeposited in the place that silver layer is not etched, after the silver layer is cut through, start etching gold
Layer, the gold atom sputtered in etching process is redeposited in the place that silver layer is not etched and etched formed nano-pore
Hole wall on, until place and the gold atom that is sputtered out of hole wall of etched formed nano-pore that silver layer is not etched are complete
Covering and nano-pore vertical depth are less than the sum of silver layer and the thickness of layer gold, obtained composition metal nanohole array substrate;
(2) needle point is prepared
After using plasma washing machine, atomic force microscope probe is cleaned up, visited using magnetron sputtering or vacuum evaporation
Wire surface deposits silverskin, then using focused-ion-beam lithography technology, using helium source, selects 15 μm of diaphragms, is 1pA in line,
Focused ion beam dose is 0.01-0.02nC/ μm2Under the conditions of, probe tip is etched, adjusts the radius of curvature of needle point, and in needle
Sharp surface etch striped.
9. a kind of local Electromagnetic enhancement device for Raman Characterization of claim 1-6 any one of them is in unimolecule
Application in Raman fingerprint characterization.
10. a kind of use of local Electromagnetic enhancement device for Raman Characterization of claim 1-6 any one of them
Method, which is characterized in that the method is specially:By sample to be tested drop in the compound gold of claim 1-6 any one of them
Belong on nanohole array substrate, then utilize simultaneously and claim 1-6 any one of them atomic force microscope probes are installed
Atomic force microscope and Raman common focusing micro spectrum instrument composition metal nanohole array substrate surface is scanned, compare
It analyzes on composition metal nanohole array substrate in nano-pore, between two adjacent nano holes, between three adjacent nano holes
Needle point enhances scattered signal feature peak position and peak intensity, obtains difference of the single testing molecule in substrate surface configuration conformation.
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CN109142796A (en) * | 2018-09-04 | 2019-01-04 | 南京航空航天大学 | A kind of object sunk area surface geometry appearance restoring method and system |
CN109142796B (en) * | 2018-09-04 | 2019-11-15 | 南京航空航天大学 | A kind of object sunk area surface geometry appearance restoring method and system |
CN109991208A (en) * | 2019-05-05 | 2019-07-09 | 中国科学院重庆绿色智能技术研究院 | Blue-green alge Species Competition allelopathy mechanism research method based on Surface Enhanced Raman Scattering Spectrum |
CN109991208B (en) * | 2019-05-05 | 2021-06-15 | 中国科学院重庆绿色智能技术研究院 | Blue-green algae population competition allelopathy mechanism research method based on surface enhanced Raman scattering spectrum |
CN110628601A (en) * | 2019-10-19 | 2019-12-31 | 上海新微技术研发中心有限公司 | Flexible substrate nanopore structure and nanopore array manufacturing method |
CN110628601B (en) * | 2019-10-19 | 2023-06-30 | 上海新微技术研发中心有限公司 | Flexible substrate nanopore structure and nanopore array manufacturing method |
CN111157509A (en) * | 2019-12-31 | 2020-05-15 | 阜阳师范大学 | Suspended substrate for enhancing Raman effect and preparation method thereof |
CN111650177A (en) * | 2020-05-09 | 2020-09-11 | 浙江大学 | Gallium telluride based surface enhanced Raman substrate for realizing monomolecular detection and preparation method thereof |
CN111650177B (en) * | 2020-05-09 | 2024-01-09 | 浙江大学 | Gallium telluride-based surface enhanced Raman substrate for realizing single-molecule detection and preparation method thereof |
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