CN105259157B - A kind of focus visible surface enhanced Raman scattering substrate, preparation method and the method using the substrate detection molecules - Google Patents
A kind of focus visible surface enhanced Raman scattering substrate, preparation method and the method using the substrate detection molecules Download PDFInfo
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Abstract
The present invention relates to a kind of visible surface enhanced Raman scattering substrate of focus, preparation method and the method using the substrate detection molecules.SERS substrates are made up of the nano silver wire being placed on Raman scattering substrate and the gold nano grain modified on nano silver wire surface, are formd plasma resonance in substrate between nano silver wire and gold nano grain, are generated strong electromagnetic field so as to form visible focus.SERS substrates are placed under Raman light microscope, find the position i.e. SERS focuses of nano silver wire surface modification gold nano grain, by testing molecule slowly be added drop-wise to the position, testing molecule is effectively dropped into SERS hotspot ranges.Preparation method simply utilizes capillary force and Action of Gravity Field can by gold nano particle modification to nano silver wire surface.The substrate SERS " focus is visible " of preparation, can effectively be such that testing molecule drops into the range of " focus ", will directly improve the sensitivity of testing result.
Description
Technical field
The present invention relates to SERS technology and the detection method of correlation, being specifically related to a kind of focus can
Surface enhanced Raman scattering substrate, preparation method and the method using the substrate detection molecules seen.
Background technology
SERS (SERS) phenomenon is found from Martin Fleischmann in 1974 and its partner, is arrived
1977, Jeanmaire and Van two scientists of Duyne are surprisingly found that to this give theoretical explanation after, SERS skills
Art is developed so far the history of nearly 40 years.SERS technologies, because it provides molecule abundant finger print information, it can distinguish same
The architectural difference of enantiomers, different orientation of the identical testing molecule in SERS substrates can be distinguished, is realized highly sensitive
Detection, therefore widely studied and applied, it is a kind of advanced analyzing detecting method.
The development of SERS technologies, between the development of strong dependences substrate, only noble metal (golden or silver etc.) substrate
Plasma resonance, which produces strong-electromagnetic field, just can amplify the Raman signal of molecule.Research shows, determines obtained molecule absorption peak
Integrated intensity it is directly proportional to the biquadratic of electromagnetic field.Therefore in existing SERS researchs, it is most of to be all directed to research regulation and control
The gap of noble metal nano particles, in particular up to gap are less than less than 10 nanometers, produce most strong plasma resonance, are formed high
The SERS " focus " of effect.
But SERS is a kind of surface spectral technique, only when testing molecule fall the surface of metallic substrates or near,
The Raman signal of molecule can be just enhanced, and this is a main feature of SERS technologies application.In existing research, most of base
The characteristics of bottom shows be：Blindly technologies such as " focuses " is looked for all to cause SERS testing results when " focus " skewness, detection
Enhancer Distribution value and its wide in range, relative average debiation is larger, cannot also meet highly sensitive qualitative analysis and
Quantitative analysis requirement.
The content of the invention
The technical problem to be solved in the present invention is a kind of visible surface enhanced Raman scattering substrate of focus of offer and system
Preparation Method, while overcome the deficiency that existing SERS substrates blindly find " focus " technology, there is provided " focus can by a kind of single SRES
See ", the SERS detection methods that make testing molecule effectively drop into the range of substrate " focus " (utilize the substrate detection molecules
Method).
In order to solve the above-mentioned technical problem, the present invention is achieved by the following technical programs：
A kind of visible surface enhanced Raman scattering substrate of focus, by the nano silver wire that is placed on Raman scattering substrate and
Modify the gold nano grain on nano silver wire surface to form, plasma is formd between nano silver wire and gold nano grain in substrate
Resonance, generates strong electromagnetic field so as to form visible focus；The length of nano silver wire be 5~15 μm, nano silver wire it is straight
Footpath is 50~150nm, a diameter of 50~150nm of gold nano grain.
A kind of preparation method of the visible surface enhanced Raman scattering substrate of focus, first synthesis of silver nano-wire and gold nano
Two kinds of colloidal sols of particle are simultaneously diluted to identical multiple, take nano silver wire colloidal sol to drop on Raman scattering substrate and dry, finally take
Gold nano particle colloidal sols drop to same area on Raman scattering substrate, in the effect of the capillary force and gravity of gas-liquid interface
Under, gold nano grain is modified nano silver wire surface, produces the visible surface enhanced Raman scattering substrate of focus.
Preferably, the preparation method of nano silver wire is：10mL ethylene glycol is added into reaction vessel, it is added at 160 DEG C
Heat backflow 2h, then injection simultaneously is dissolved in 0.1M silver nitrate solutiones in 5mL ethylene glycol solvents, molten thereto with two syringe pumps
0.222g polyvinylpyrrolidone of the solution in 5mL ethylene glycol solvents, charge velocity 0.2mL/min；Finally, in 160 DEG C of perseverances
The lower reaction 60min of temperature, the nano silver wire colloidal sol for synthesizing to obtain is according to the ratio that volume ratio is 1: 9, successively with acetone, secondary washing
Wash twice.
Preferably, the preparation method of gold nano grain is：90mL secondary water, 1mL mass point is added into reaction vessel
Number is 1% chlorauric acid solution, stirs, is heated to seething with excitement, and is rapidly added the sodium citrate solution that 1mL mass fractions are 1% and makees
For reducing agent, gold nano gold seeds is obtained；Gold nano gold seeds 10mL is taken, is separately added into the hydrochloric acid hydroxyl that concentration is 25mM thereto
Amine 0.4mL, the sodium citrate solution 0.4mL and secondary water 28.4mL that mass fraction is 1%, stir 5min, finally add
Mass fraction is 1% gold chloride 0.8mL, at room temperature stirring reaction 1h；It is 1 that obtained gold nano grain, which is synthesized, according to volume ratio
: 10 ratio, with secondary water washing three times.
Preferably, the preparation method of SERS substrates is：By the silicon chip soaked in advance with chloroazotic acid respectively with running water, secondary
After water, ethanol washing three times, dried up with nitrogen；Nano silver wire is diluted with water according to 1: 100 ratio, 1 μ is pipetted with liquid-transfering gun
L is added drop-wise on silicon chip；After drying, gold nano colloidal sol is taken, after same 100 times of dilution, 1 μ L is pipetted with liquid-transfering gun and is added drop-wise to the silicon chip
On, with the volatilization of solvent, in the presence of the capillary force and gravity of gas-liquid interface, gold nano grain is modified silver nanoparticle
Line surface, obtain the visible surface enhanced Raman scattering substrate of focus.
Using the method for the visible surface enhanced Raman scattering substrate detection molecules of focus, by SERS
(SERS) substrate is placed under Raman light microscope, finds the i.e. SERS in the position heat of nano silver wire surface modification gold nano grain
Point, by testing molecule slowly be added drop-wise to the position, testing molecule is effectively dropped into SERS hotspot ranges, detection signal
Measure obtains the SERS spectrograms of the molecule.
The visible surface enhanced Raman scattering substrate of focus of the present invention, preparation method and using the substrate detection molecules
Method, the analysis of its principles of science：
First, gold nano grain similar in diameter and nano silver wire is arrived into nano silver wire by capillary force and gravity modification
Behind surface, plasma resonance is formed between two kinds of particles, can be to produce strong electromagnetic field, i.e. SERS " focus ".
2nd, length is about 10 microns of nano silver wire, and its longitudinal absworption peak has been moved to region of ultra-red, conventionally used
Raman excitation light action under, it is impossible to form good resonance Raman, therefore the excitation wavelength selection to measure does not interfere with.
3rd, during Raman detection, micron-sized nano silver wire can be modified in its table by directly being observed under light microscope
The gold nano grain in face, be contrasted with direct similar nano silver wire, thus its be distributed in the position of nano silver wire can be in light
Learn and narrow down to a certain scope under microscope, and this scope is smaller than the laser-formed hot spot scope used in Raman detection, thus it is silver-colored
" focus " formed between nano wire and gold nano grain can be controlled in the range of hot spot, realize " visible " operation, so as to
Molecule is effectively added drop-wise in the range of " focus ".
Relative to prior art, beneficial effects of the present invention performance is as follows：
1), conventional art synthesis SERS substrates in, typically will utilize specific molecular connect two kinds of noble metals, and these
Connection molecule is likely to have signal interference when SERS is detected, and the preparation method of the present invention need not add connection point in addition
Son, capillary force and Action of Gravity Field can are simply utilized by gold nano particle modification to nano silver wire surface.
2) it is most of all quickly, accurately to find " focus ", in existing SERS substrates, the homogeneity of whole substrate
Difference have impact on the reappearance of result, and substrate SERS " focus is visible " prepared by the present invention, effectively can fall testing molecule
Enter in the range of " focus ", will directly improve the sensitivity of testing result.
3), the method for preparation method of the invention and detection molecules is simple, has wide market application foreground.
Brief description of the drawings
Fig. 1 be the present invention SERS substrates in single gold nano particle modification to nano silver wire surface schematic diagram and reality
Dark field microscope compares figure before and after the nano silver wire repairing gilding nano particle on border.
Fig. 2 be the SERS substrate Raman optical microscopes that prepare of the present invention and detection 4-ATP molecules SERS spectrograms and
Region SERS mapping scheme.
The SERS substrates that Fig. 3 is the present invention are used for the cysteine of low concentration, atriphos, crystal methamphetamine and often
See the SERS detection spectrograms of agricultural chemicals paraoxon.
Embodiment
The preferred embodiment of the present invention is described in further detail below in conjunction with the accompanying drawings.
Embodiment 1
Step 1: the preparation of nano silver wire
10mL ethylene glycol is added into 100mL three-necked flasks, it is heated to reflux to 2h at 160 DEG C, then with two syringe pumps
0.1M silver nitrate solutiones, 0.222g polyvinylpyrrolidones (being dissolved in respectively in 5mL ethylene glycol solvents) are injected simultaneously thereto,
Charge velocity is 0.2mL/min.Finally, 60min is reacted under 160 DEG C of constant temperature, the nano silver wire colloidal sol for synthesizing to obtain is according to volume
Than the ratio for 1: 9, successively washed twice with acetone, secondary water, obtained nano silver wire length is 10 μm, diameter is about
110nm。
Step 2: the preparation of gold nano grain：
90mL secondary water is added into 500mL three-necked flasks, the chlorauric acid solution that 1mL mass fractions are 1%, magnetic force stirs
Mix, oil bath heating to seethe with excitement after, be rapidly added 1mL mass fractions be 1% sodium citrate solution as reducing agent, obtained gold
Nano-particle diameter is about 50nm, and the gold seeds that large-size particle is obtained as further growth is standby.
A diameter of 50nm obtained above aurosol 10mL is taken, is separately added into the hydroxylamine hydrochloride that concentration is 25mM thereto
0.4mL, the sodium citrate solution 0.4mL and secondary water 28.4mL that mass fraction is 1%, magnetic agitation 5min, finally add again
Enter the gold chloride 0.8mL that mass fraction is 1%, at room temperature stirring reaction 1h.Obtained gold nano grain is synthesized according to volume ratio
For 1: 10 ratio, with secondary water washing three times, the diameter of obtained gold nano grain is about 100nm.
Step 3: the preparation of SERS substrates：
After silicon chip after soaking 24h with chloroazotic acid in advance is washed three times with running water, secondary water, ethanol respectively, nitrogen is used
Drying.The nano silver wire that step 1 synthesizes is diluted with water according to 1: 100 ratio, pipetting 1 μ L with liquid-transfering gun is added drop-wise to silicon chip
On；After 1min or so is dried, the gold nano colloidal sol of the synthesis of above-mentioned steps two is taken, after being equally diluted with water 100 times, is moved with liquid-transfering gun
1 μ L are taken to be added drop-wise on the silicon chip, with the volatilization of solvent, in the presence of the capillary force and gravity of gas-liquid interface, gold nano
Particle is modified nano silver wire surface, obtains the SERS substrates of " visible hot spots " that the present invention synthesizes.
Can be seen that the evaporation with solvent from Figure 1A, B, the capillary force and gravity of gas-liquid interface by a few
Gold nano particle modification is near nano silver wire or surface.Fig. 1 C, D shoot for details in a play not acted out on stage, but told through dialogues, correspond to nano silver wire and are modifying
Contrast photo before and after gold nano grain, the signified i.e. modification of arrow is in the gold nano grain position on nano silver wire surface in Fig. 1 D.
Embodiment 2
4-ATP molecules are determined using the SERS substrates of the invention prepared, obtain SERS spectrograms and region SERS
Mapping images：
The SERS substrates that embodiment 1 synthesizes are placed under Raman light microscope, find nano silver wire surface modification
The position of gold nano grain, that is, SERS " focus " position is produced, 1 μ L concentration is pipetted as 10 with liquid-transfering gun-8M 4-ATP is molten
Liquid, slowly be added drop-wise to " focus " position or near.After 1min or so is dried, selection wavelength is 633nm, laser power is
2mW exciting light, 1 second time of integration, detection signal, obtain the SERS spectrograms of 4-ATP molecules.
Fig. 2A is the SERS substrate Raman optical microscopes that prepare of the present invention, wherein square frame mark for raman laser
Hot spot scope：(4 × 4) square micron；Fig. 2 B are schematic diagram when substrate prepared by the present invention is used to determine Molecular Raman signal：
Wherein single SERS " focus " and testing molecule effectively fall in the range of hot spot；Fig. 2 C are the substrate prepared using the present invention
The SERS spectrograms of obtained 4-ATP molecules are determined, the wherein color of ordinate is used for showing the integration of the characteristic peak of 4-ATP molecules
Intensity, black, green, red, yellow represent gradually to strengthen.
Meanwhile selection wavelength is 633nm, the exciting light that laser power is 2 milliwatts, scanning stepping is 0.5 μm/point, with
Centered on " visible hot spots ", scan area is 4 × 4 μm2, finally with 1438cm-14-ATP characteristic peak integrated intensity values formed
SERS mapping images.Fig. 2 D are the region SERS that the substrate prepared with the present invention determines obtained 4-ATP molecules
Mapping schemes (1438cm-1The 4-ATP characteristic peaks at place), the color in region equally represents that the signal integration of characteristic peak herein is strong
Degree change, represent gradually to strengthen from black to yellow, clearly show " focus " scope and change.
Embodiment 3
Cysteine, atriphos, crystal methamphetamine and to oxygen are determined using the SERS substrates that prepare of the present invention
Phosphorus：
With the identical method of above-described embodiment 2, concentration can be respectively obtained as 10-6M cysteine, 10-6M three phosphorus
Adenosine monophosphate, 10-7M crystal methamphetamine and 10-8The SERS spectrograms of M paraoxon.
In Fig. 3, A：10-6M cysteine, B：10-6M atriphos, C：10-7M crystal methamphetamine, D：10-8M
Paraoxon, as a result show cysteine common in either human body and atriphos molecule, or common methamphetamine methyl
Phenylpropyl alcohol amine molecule and agricultural chemicals paraoxon molecule, the SERS substrates prepared using the present invention, can reach high sensitivity SERS
Detection results.
For the various embodiments described above simply to illustrate that the technical concepts and features of the present invention, the purpose is to be to allow in the art
Those of ordinary skill can understand present disclosure and implement according to this, it is not intended to limit the scope of the present invention.
Equivalent change or modification made by every essence according to present invention, should all cover in protection scope of the present invention
It is interior.
Claims (2)
1. a kind of visible surface enhanced Raman scattering substrate of focus, by the nano silver wire being placed on Raman scattering substrate and is repaiied
Adorn the gold nano grain on nano silver wire surface to form, foring plasma between nano silver wire and gold nano grain in substrate is total to
Shake, generate strong electromagnetic field so as to form visible focus；The length of nano silver wire is 5~15 μm, the diameter of nano silver wire
For 50~150nm, a diameter of 50~150nm of gold nano grain；
Preparation method is：Two kinds of colloidal sols of synthesis of silver nano-wire and gold nano grain and identical multiple is diluted to first, takes Yin Na
Rice noodles colloidal sol is dropped on Raman scattering substrate and dried, and finally takes gold nano particle colloidal sols to drop to identical portions on Raman scattering substrate
Position, in the presence of the capillary force and gravity of gas-liquid interface, gold nano grain is modified nano silver wire surface, produces heat
The visible surface enhanced Raman scattering substrate of point；
In preparation method, the preparation method of nano silver wire is：10mL ethylene glycol is added into reaction vessel, it is added at 160 DEG C
Heat backflow 2h, then injection simultaneously is dissolved in 0.1M silver nitrate solutiones in 5mL ethylene glycol solvents, molten thereto with two syringe pumps
0.222g polyvinylpyrrolidone of the solution in 5mL ethylene glycol solvents, charge velocity 0.2mL/min；Finally, in 160 DEG C of perseverances
The lower reaction 60min of temperature, the nano silver wire colloidal sol for synthesizing to obtain is according to the ratio that volume ratio is 1: 9, successively with acetone, secondary washing
Wash twice；
In preparation method, the preparation method of gold nano grain is：
90mL secondary water is added into reaction vessel, the chlorauric acid solution that 1mL mass fractions are 1%, stir, be heated to boiling
Rise, be rapidly added 1mL mass fractions be 1% sodium citrate solution as reducing agent, obtain gold nano gold seeds；Take gold nano
Gold seeds 10mL, it is molten that the sodium citrate that hydroxylamine hydrochloride 0.4mL, mass fraction that concentration is 25mM are 1% is separately added into thereto
Liquid 0.4mL and secondary water 28.4mL, 5min is stirred, finally add the gold chloride 0.8mL that mass fraction is 1%, at room temperature
Stirring reaction 1h；Obtained gold nano grain is synthesized according to the ratio that volume ratio is 1: 10, with secondary water washing three times；
In preparation method, the preparation method of SERS substrates is：
After the silicon chip soaked in advance with chloroazotic acid is washed three times with running water, secondary water, ethanol respectively, dried up with nitrogen；By silver
Nano wire is diluted with water according to 1: 100 ratio, and pipetting 1 μ L with liquid-transfering gun is added drop-wise on silicon chip；After drying, take gold nano molten
Glue, after same 100 times of dilution, pipette 1 μ L with liquid-transfering gun and be added drop-wise on the silicon chip, with the volatilization of solvent, the hair of gas-liquid interface
For spy firmly and in the presence of gravity, gold nano grain is modified nano silver wire surface, obtains the visible surface enhanced of focus
Raman scattering substrate.
2. a kind of method using the visible surface enhanced Raman scattering substrate detection molecules of focus as claimed in claim 1, its
It is characterised by, SERS (SERS) substrate is placed under Raman light microscope, finds nano silver wire surface
Modify gold nano grain position be SERS focuses, by testing molecule slowly be added drop-wise to the position, make testing molecule effective
Drop into SERS hotspot ranges, detection signal determines to obtain the SERS spectrograms of the molecule.
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