CN105842225A - Silicon-based SERS chip for quantitatively detecting lead ion concentration in actual water sample and preparation method thereof - Google Patents
Silicon-based SERS chip for quantitatively detecting lead ion concentration in actual water sample and preparation method thereof Download PDFInfo
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- CN105842225A CN105842225A CN201610179491.XA CN201610179491A CN105842225A CN 105842225 A CN105842225 A CN 105842225A CN 201610179491 A CN201610179491 A CN 201610179491A CN 105842225 A CN105842225 A CN 105842225A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/65—Raman scattering
- G01N21/658—Raman scattering enhancement Raman, e.g. surface plasmons
Abstract
The invention relates to the field of environment monitoring and discloses a silicon-based SERS chip for quantitatively detecting lead ion concentration in an actual water sample and a preparation method thereof. The chip is composed of: a silicon wafer modified with silver nano particles, gold nano particles, nucleotide sequences represented as the SEQ ID No.1, 2, and 4, and a sequence fragment formed by coupling a sulfydryl group to a 5-terminal and coupling a fluorescent dye to a 3-terminal of a nucleotide sequence SEQ ID No.3. In the invention, the surface enhanced Raman scattering silicon-based chip with assistance of polyadenine is employed for detecting the lead ion, wherein the chip is composed of core (silver) and satellite (gold) nano particles, wherein the DNAzyme covalently connected to the chip can be excited specifically by the lead ions, so that the substrate strand is broken into two free DNA segments. The chip can detect a strong SERS signal, and achieves high sensitivity, specificity, repeatability and recyclability.
Description
Technical field
The present invention relates to environmental monitoring technology field, be specifically related to a kind of detection by quantitative actual water sample
Silica-based SERS chip of middle plumbum ion concentration and preparation method thereof.
Background technology
Lead ion is one of heavy metal ion the most dangerous in environment, there is multiple source and such as eats
Thing, blood, drinking water, industrial wastewater etc..Research finds, by food chain and drinking water
The nervous system of the mankind particularly child, urinary system, intelligent development etc. can be caused by lead contamination
Serious injury (sees: Int.J.Hyg.Environ.Health 2008,211,345-351；Annu.
Rev.Med.2004,55,209-222).Lead ion is not easy to degrade in human body, thus for a long time
The most a small amount of lead ion of accumulation also can affect the health of the mankind.Environmental Protection Agency USA specifies
Maximum residual volume at Pb in Drinking Water ion not can exceed that 72.4nM.Have been reported discovery, one
A little children expose with low-level lead ion due to long-term, and mental capacity is affected by serious
(seeing: British Medical Bulletin 2003,68,167-182).Therefore for lead ion
Detection be increasingly becoming the major issue of environmental monitoring, research and develop NEW Pb ion detection method pair
Significant in environmental conservation, disease prevention, great environmental pollution monitoring.
At present, various traditional analysis methods are as atomic absorption spectrography (AAS), inductively coupled plasma
Body-mass spectrography, Inductive coupling plasma-atomic emission spectrometry, it is widely used.But,
These analysis methods are expensive mostly, time-consuming, need tediously long step and complicated instrument,
Thus limit the extensive application of lead ion detection.
Surface enhanced raman spectroscopy (SERS) is the most attracting phenomenon of one, and it is the most extensive
In Application in Sensing.Comparing more normal Raman signal, Molecular Adsorption is specific at some
During metal surface, the Raman signal of this molecule can be greatly improved, it is thus possible to superelevation spirit
Quick detection sample.This huge enhancer be due to, under light illumination, for surface relatively
For coarse metal substrate, surface can produce local electromagnetic field.Except higher sensitive
Property, SERS has narrow Raman peaks thus causes less background, beneficially multivariate detection.
Additionally, Raman scattering is the most highly stable, it is little affected by humidity, oxygen, outward
Carry out the impact of species etc..Therefore, SERS has been used for susceptiveness and specific detection lead ion.
Such as, king et al., develop using with simple " Signal off " SERS strategy of a kind of uniqueness
In susceptiveness and optionally detect lead ion (see: Chem.Commun.2011,47,
4394-4396).Afterwards, Xu et al., describe a kind of Ag NPs-on-Ag based on DNAzyme
Film structure, it is possible to detection 1nM lead ion (see: Anal.Chem.2014,86,
11494-11497).Although these SERS sensors are feasible, but susceptiveness and repeatability
It not gratifying, thus limit its application in actual water sample.Therefore, more exert
Power starts to be devoted to development stability is good, repeatability is excellent substrate to realize ultrasensitiveness and spy
The detection product of the opposite sex.
Summary of the invention
In view of this, it is an object of the invention to provide in a kind of detection by quantitative actual water sample lead from
Silica-based SERS chip of sub-concentration and preparation method thereof so that described silica-based SERS chip is in inspection
Have when surveying plumbum ion concentration in actual water sample preferable sensitivity, specificity, repeatability and
Recyclability.
For achieving the above object, the present invention provides following technical scheme:
The silica-based SERS chip of plumbum ion concentration, its feature in a kind of detection by quantitative actual water sample
Be, by the silicon wafer of modified by silver nanoparticles, gold nano grain, SEQ ID NO:1-2 and
Nucleotide sequence shown in SEQ ID NO:4, and at nucleotide sequence 5 ' shown in SEQ ID NO:3
End coupling has sulfydryl, 3 ' end couplings to have the sequence fragment of fluorescent dye；
Wherein, the silicon wafer of modified by silver nanoparticles and nucleotide sequence shown in SEQ ID NO:1
Connecting, gold nano grain is connected with nucleotide sequence shown in SEQ ID NO:2, SEQ ID
Nucleotide sequence shown in NO:1-2 forms complementary double-strand each other and connects, shown in SEQ ID NO:3
Nucleotide sequence 5 ' end coupling has sulfydryl, 3 ' end couplings to have the sequence fragment of fluorescent dye by end
End sulfydryl covalently bound with gold, silver nano-particle, nucleotide sequence shown in SEQ ID NO:4 and
Nucleotide sequence shown in SEQ ID NO:3 forms complementary double-strand each other and connects.
The present invention is directed to the SERS sensitivity of plumbum ion concentration in existing detection actual water sample
The problem that (only nM level) and repeatability are poor, the present invention uses poly adenine (Poly
Nucleotide sequence shown in A30, i.e. SEQ ID NO:1-2, is respectively designated as Poly in the present invention
A30-P1 and Poly A30-P2) the surface enhanced raman spectroscopy silicon base chip that assists is for high-performance
Lead ion detection, this chip of the present invention is nano-particle modified by core (silver-colored)-satellite (golden)
Silicon wafer constituted, covalently bound DNAzyme (i.e. SEQ on silica-based SERS chip
Nucleotide sequence shown in ID NO:3, named Cy5-17E-SH in the present invention) can by lead from
Sub-specific activation so that substrate strand (i.e. nucleotide sequence shown in SEQ ID NO:4,
Named 17DS in the present invention) fragment into two sections of DNA freely, thus can detect that stronger
SERS signal, principle schematic is shown in Fig. 1.
Wherein, as preferably, described sequence fragment is at nucleotides sequence shown in SEQ ID NO:3
Row 5 ' end coupling has HS-(CH2)6-, 3 ' end couplings have a sequence fragment of Cy5 fluorescent dye, i.e. 5 '
-HS-(CH2)6-TTTCATCTCTTCTCCGAGCCGGTCGAAATAGTGAGT-Cy
5-3’。
As preferably, described silicon wafer is p-type or the n-type silicon wafer of 0.01～20 Ω * cm.
Meanwhile, present invention also offers the preparation method of described silica-based SERS chip, including:
Step 1, the silicon wafer preparing modified by silver nanoparticles and gold nano grain；
Step 2, the silicon wafer of the modified by silver nanoparticles prepared is delayed with being dissolved in phosphate
Rush the mixing of nucleotide sequence constant temperature shown in the SEQ ID NO:1 in liquid to hatch, add the most wherein
Enter saline solution, the most aging, obtain the Yin Na that nucleotide sequence shown in SEQ ID NO:1 connects
The silicon wafer that rice grain is modified；
By prepared gold nano grain and the SEQ ID NO:2 being dissolved in phosphate buffer
Shown nucleotide sequence constant temperature mixing is hatched, and then adds saline solution in solution, the most aging,
Obtain the gold nano grain that nucleotide sequence shown in SEQ ID NO:2 connects；
Step 3, step 2 gained silicon wafer and gold nano grain are placed in constant temperature in hybridization buffer
Mixing is hatched, and forms double-strand by the pairing of nucleotide sequence complementary shown in SEQ ID NO:1-2 and ties
Structure, rinses with PBS, and then nitrogen dries up, and obtains core (silver-colored)-satellite (golden) nano-particle
The silicon wafer (scanning electron microscope characterizes photo and sees Fig. 2) modified；
Step 4, by silicon wafer nano-particle modified for core (silver-colored)-satellite (golden) with at SEQ ID
The end coupling of nucleotide sequence 5 ' shown in NO:3 has sulfydryl, 3 ' end couplings to have the tract of fluorescent dye
Section constant temperature blending oscillating reactions, make nucleotide sequence terminal sulfhydryl group shown in SEQ ID NO:3 with
Gold, silver nanoparticle covalently bound formation gold-sulfide linkage and silver-sulfide linkage, then add salt in solution
Solution is the most aging；
Material after aging is taken out, and is dissolved in hybridization buffer shown in SEQ ID NO:4
The mixing of nucleotide sequence constant temperature is hatched, and forms DNA double chain structure by DNA complementary pairing,
Then rinsing with PBS, nitrogen dries up, and obtains described silica-based SERS chip.
As preferably, the silicon wafer of described modified by silver nanoparticles is prepared by following methods:
Monocrystalline silicon piece is carried out ultrasonic cleaning, so with deionized water, acetone, deionized water successively
After clean with concentrated sulphuric acid and mixed solution of hydrogen peroxide again；
Monocrystalline silicon piece after cleaning joins and carries out silicon-hydrogenation in hydrofluoric acid solution, obtains table
Face covers the silicon wafer of Si--H bond, and then light faces up, and puts into the mixing of silver nitrate and Fluohydric acid.
In solution, slow oscillating reactions, silver ion is reduced by Si--H bond, the most raw at silicon wafer surface
The uniform silver nano-grain of long last layer, obtains the silicon wafer of modified by silver nanoparticles, finally uses
Nitrogen dries up surface.
As preferably, described hydrogen peroxide mass concentration is 40%, concentrated sulphuric acid and 40% peroxide
Change hydrogen volume ratio for 1:(0.01～100).
As preferably, in described hydrofluoric acid solution, the mass concentration of Fluohydric acid. is 1～40%.
As preferably, the time of described silicon-hydrogenation is 1～60 minute.
As preferably, the mixed solution of described silver nitrate and Fluohydric acid. is by the silver nitrate solution of 1M
With the hydrofluoric acid solution that mass concentration is 40% by volume for 1:(0.01～100) formulated.
As preferably, the described oscillating reactions time is 1～60 minute.
As preferably, described nanogold particle is prepared by citric acid reducing process.Concrete
Can refer to following manner:
In the chlorauric acid solution of boiling, add sodium citrate solution, after stirring reaction,
To gold nano grain, described gold chloride mass concentration is 0.01%, and sodium citrate mass concentration is
1%, the response time is 15 minutes.
As preferably, nucleotide sequence concentration shown in SEQ ID NO:1-2 is 0.001～1M.
As preferably, constant temperature mixing described in step 2 is hatched as hatching 16 hours at 25 DEG C.
As preferably, add saline solution described in step 2 and step 4 for being 1M by initial concentration
Saline solution, add every two little time-division 3-5 time, saline solution ultimate density is 0.01～1M.
As preferably, constant temperature mixing described in step 3 is hatched as hatching 24 hours at 37 DEG C.
As preferably, oscillating reactions described in step 4 be 100～600 rpms, at 25 DEG C
React 1～24 hour.
As preferably, constant temperature mixing described in step 4 is hatched as hatching 1～24 hour at 37 DEG C.
As preferably, step 4 has in the end coupling of nucleotide sequence 5 ' shown in SEQ ID NO:3
Sulfydryl, 3 ' end couplings have nucleotide sequence shown in the sequence fragment of fluorescent dye, SEQ ID NO:4
Concentration is 0.001～1M.
The silica-based SERS chip that the present invention builds is from the log concentration of 10pM to 1 μM of lead ion
Preferable linear relationship (R is there is with normalized raman scattering intensity2=0.997)；Owing to this chip is excellent
SERS performance more, it can detect as little as 8.9 × 10-12The lead ion of M (pM level) is dense
Degree, well below the SERS sensor (nM level) reported；Additionally, the chip provided
There is good selectivity and recyclability (after three circulations, Raman loss of strength is only 11.1%)；
The more important thing is, prepared chip can accurately and reliably detect lake water, tap water and work
The concentration (RSD value is less than 12%) of unknown lead ion in the practical systems such as industry waste water.
Accompanying drawing explanation
Fig. 1 is the schematic diagram of the silica-based SERS chip detection lead ion of the present invention；
Fig. 2 is the silica-based SERS chip that the present invention prepares, i.e. core (silver-colored)-satellite (golden)
The scanning electron microscope of nano-particle modified silicon wafer characterizes photo, wherein silver nano-grain (Ag
NPs) size is about 110nm, and the size of gold nano grain (Au NPs) is about 13nm,
And more uniform be distributed on Ag NPs surface；
Fig. 3 is that the SERS of variable concentrations lead ion is tested system by the chip that the present invention prepares
Row result spectrogram；
Fig. 4 is the chip for preparing of the present invention SERS spectra to same concentration different ions
Figure and Raman peaks 1366cm of different ions-1The contrast bar diagram of intensity, wherein a figure is
SERS spectra figure to same concentration different ions, b figure is the Raman peaks of different ions
1366cm-1The contrast bar diagram of intensity；
Fig. 5 is the schematic diagram of the chip reconstruction that the present invention prepares and recycles for 3 times
SERS spectra figure and corresponding Raman peaks 1366cm-1The comparison diagram of intensity, wherein a figure is core
The reconstruction schematic diagram of sheet, b and c figure is respectively the SERS spectra figure recycled 3 times with corresponding
Raman peaks 1366cm-1The comparison diagram of intensity.
Detailed description of the invention:
The invention discloses the silica-based SERS of plumbum ion concentration in a kind of detection by quantitative actual water sample
Chip and preparation method thereof, those skilled in the art can use for reference present disclosure, be suitably modified work
Skill parameter realizes.Special needs to be pointed out is, all similar replacements and change are to this area skill
Being apparent from for art personnel, they are considered as being included in the present invention.The product of the present invention
Product and method are described by preferred embodiment, related personnel substantially can without departing from
Method described herein and application are modified in present invention, spirit and scope or suitably
Change and combination, realize and apply the technology of the present invention.
Plumbum ion concentration silica-based in a kind of detection by quantitative the actual water sample below present invention provided
SERS chip and preparation method thereof is described further.
Embodiment 1: prepare silica-based SERS chip of the present invention
(1) Fluohydric acid. auxiliary etch method prepares the silicon wafer of modified by silver nanoparticles
Take 0.5cm2Size monocrystalline silicon piece 3-6 sheet is placed in clean beaker and spends successively in Ultrasound Instrument
Ionized water, acetone, deionized water carry out ultrasonic cleaning 15 minutes, place into 40mL concentrated sulphuric acid and
Hydrogen peroxide (mass concentration: 40%) mixed solution (volume ratio=3:1) cleans further,
Clean with deionized water the most again, obtain clean silicon wafer.The silicon wafer cleaned up is put
Enter in hydrofluoric acid solution (mass concentration: 5%) and carry out silicon-hydrogenation, slowly vibration 30 minutes,
Obtain surface and cover the silicon wafer of a large amount of Si--H bond.By silicon wafer obtained after above-mentioned process
Sheet, light faces up, and puts into 20mL silver nitrate (1M) and Fluohydric acid. (mass concentration: 40%)
Mixed solution (volume ratio=1:100) in, slow oscillating reactions 60 minutes, anti-according to electrochemistry
Principle, silver ion is answered to be reduced by Si--H bond, uniform at silicon wafer surface growth in situ last layer
Silver nano-grain, thus obtain the silicon wafer of modified by silver nanoparticles, finally dry up table with nitrogen
Face.
(2) preparation of the silicon wafer that core (silver-colored)-satellite (golden) is nano-particle modified
Citric acid method of reducing according to standard, at the 50mL chlorauric acid solution seethed with excitement, (quality is dense
Degree: 0.01%) in, add 2mL sodium citrate solution (mass concentration: 1%), stirring reaction
After 15 minutes, obtain gold nano grain.By prepared gold nano grain be dissolved in phosphoric acid
Poly A30-P1DNA (5 ' in salt buffer (PBS)
-AAAAAAAAAAAAAAAAAAAAAAAAAAAAAATTTTTGATACAG
CTAATTCAGAATCATTTTGTGGA-3 ') (concentration: 0.001M), constant temperature 25 DEG C,
On constant temperature blending instrument, mixing is hatched 16 hours, then divides in solution and added one every 2 hours 3 times
Secondary 1M saline solution so that saline solution ultimate density is 0.1M, the most aging, obtains Poly
The gold nano grain that A30-P1 connects.By the silicon wafer of modified by silver nanoparticles that prepared with
The Poly A30-P2DNA (5 ' being dissolved in phosphate buffer
-AAAAAAAAAAAAAAAAAAAAAAAAAAAAAATTTTTTCCACAA
AATGATTCTGAATTAGCTGTATC-3 ') (concentration: 0.001M), constant temperature 25 DEG C,
On constant temperature blending instrument, mixing is hatched 16 hours, then divides in solution and added one every 2 hours 5 times
Secondary 1M saline solution so that saline solution ultimate density is 0.1M, the most aging, obtains Poly
The silicon wafer of the modified by silver nanoparticles that A30-P2 connects.Above-mentioned two step resulting materials are placed in miscellaneous
Handing in buffer, constant temperature 37 DEG C, on constant temperature blending instrument, mixing hatches 24 hours.Pass through DNA
Complementary pairing forms duplex structure, rinses several times with PBS, and then nitrogen dries up, and obtains core (silver-colored)
The silicon wafer that-satellite (golden) is nano-particle modified.
(3) the SERS chip structure of the silicon wafer that core (silver-colored)-satellite (golden) is nano-particle modified
Build
Silicon wafer nano-particle modified for core (silver-colored)-satellite (golden) is placed in centrifuge tube, adds
Enter enzyme strand DNA (Cy5-17E-SH, 5 '
-HS-(CH2)6-TTTCATCTCTTCTCCGAGCCGGTCGAAATAGTGAGT-C
Y5-3 ') solution of (concentration: 0.001M) so that solution submergence material.Centrifuge tube is put
In constant temperature blending instrument, 350 rpms, 25 DEG C of constant temperature, oscillating reactions 24 hours, make DNA
Terminal sulfhydryl group and gold, silver nanoparticle covalency form gold-sulfide linkage and silver-sulfide linkage, make DNA covalency even
Receive on the silicon wafer that core (silver-colored)-satellite (golden) is nano-particle modified.Then in solution, divide 5
Secondary added a 1M saline solution every 2 hours so that saline solution ultimate density is 0.1M, the most always
Change.By material take out, be dissolved in substrate strand DNA in hybridization buffer (17DS,
5’-ACTCACTATrAGGAAGAGATG-3’,rA(ribonucleotide adenosine)
Represent base A in ribonucleotide) (concentration: 0.001M), 37 DEG C of constant temperature, at constant temperature
In blending instrument, mixing is hatched 24 hours, forms DNA double chain structure by DNA complementary pairing, so
Rinsing several times with PBS afterwards, nitrogen dries up, and obtains core (silver-colored)-satellite (golden) nano-particle and repaiies
The SERS chip of the silicon wafer of decorations.
Embodiment 2: prepare silica-based SERS chip of the present invention
(1) Fluohydric acid. auxiliary etch method prepares the silicon wafer of modified by silver nanoparticles
Take 0.5cm2Size monocrystalline silicon piece 3-6 sheet is placed in clean beaker and spends successively in Ultrasound Instrument
Ionized water, acetone, deionized water carry out ultrasonic cleaning 15 minutes, place into 40mL concentrated sulphuric acid and
Hydrogen peroxide (mass concentration: 40%) mixed solution (volume ratio=3:1) cleans further,
Clean with deionized water the most again, obtain clean silicon wafer.The silicon wafer cleaned up is put
Enter in hydrofluoric acid solution (mass concentration: 5%) and carry out silicon-hydrogenation, slowly vibration 30 minutes,
Obtain surface and cover the silicon wafer of a large amount of Si--H bond.By silicon wafer obtained after above-mentioned process
Sheet, light faces up, and puts into 20mL silver nitrate (1M) and Fluohydric acid. (mass concentration: 40%)
Mixed solution (volume ratio=1:50) in, slow oscillating reactions 30 minutes, anti-according to electrochemistry
Principle, silver ion is answered to be reduced by Si--H bond, uniform at silicon wafer surface growth in situ last layer
Silver nano-grain, thus obtain the silicon wafer of modified by silver nanoparticles, finally dry up table with nitrogen
Face.
(2) preparation of the silicon wafer that core (silver-colored)-satellite (golden) is nano-particle modified
Citric acid method of reducing according to standard, at the 50mL chlorauric acid solution seethed with excitement, (quality is dense
Degree: 0.01%) in, add 2mL sodium citrate solution (mass concentration: 1%), stirring reaction
After 15 minutes, obtain gold nano grain.By prepared gold nano grain be dissolved in phosphoric acid
Poly A30-P1DNA (5 ' in salt buffer (PBS)
-AAAAAAAAAAAAAAAAAAAAAAAAAAAAAATTTTTGATACAG
CTAATTCAGAATCATTTTGTGGA-3 ') (concentration: 0.001M), constant temperature 25 DEG C,
On constant temperature blending instrument, mixing is hatched 16 hours, then divides in solution and added one every 2 hours 3 times
Secondary 1M saline solution so that saline solution ultimate density is 0.1M, the most aging, obtains Poly
The gold nano grain that A30-P1 connects.By the silicon wafer of modified by silver nanoparticles that prepared with
The Poly A30-P2DNA (5 ' being dissolved in phosphate buffer
-AAAAAAAAAAAAAAAAAAAAAAAAAAAAAATTTTTTCCACAA
AATGATTCTGAATTAGCTGTATC-3 ') (concentration: 0.001M), constant temperature 25 DEG C,
On constant temperature blending instrument, mixing is hatched 16 hours, then divides in solution and added one every 2 hours 5 times
Secondary 1M saline solution so that saline solution ultimate density is 0.1M, the most aging, obtains Poly
The silicon wafer of the modified by silver nanoparticles that A30-P2 connects.Above-mentioned two step resulting materials are placed in miscellaneous
Handing in buffer, constant temperature 37 DEG C, on constant temperature blending instrument, mixing hatches 24 hours.Pass through DNA
Complementary pairing forms duplex structure, rinses several times with PBS, and then nitrogen dries up, and obtains core (silver-colored)
The silicon wafer that-satellite (golden) is nano-particle modified.
(3) the SERS chip structure of the silicon wafer that core (silver-colored)-satellite (golden) is nano-particle modified
Build
Silicon wafer nano-particle modified for core (silver-colored)-satellite (golden) is placed in centrifuge tube, adds
Enter enzyme strand DNA (Cy5-17E-SH, 5 '
-HS-(CH2)6-TTTCATCTCTTCTCCGAGCCGGTCGAAATAGTGAGT-C
Y5-3 ') solution of (concentration: 0.001M) so that solution submergence material.Centrifuge tube is put
In constant temperature blending instrument, 350 rpms, 25 DEG C of constant temperature, oscillating reactions 24 hours, make DNA
Terminal sulfhydryl group and gold, silver nanoparticle covalency form gold-sulfide linkage and silver-sulfide linkage, make DNA covalency even
Receive on the silicon wafer that core (silver-colored)-satellite (golden) is nano-particle modified.Then in solution, divide 5
Secondary added a 1M saline solution every 2 hours so that saline solution ultimate density is 0.1M, the most always
Change.By material take out, be dissolved in substrate strand DNA in hybridization buffer (17DS,
5’-ACTCACTATrAGGAAGAGATG-3’,rA(ribonucleotide adenosine)
Represent base A in ribonucleotide) (concentration: 0.001M), 37 DEG C of constant temperature, at constant temperature
In blending instrument, mixing is hatched 24 hours, forms DNA double chain structure by DNA complementary pairing, so
Rinsing several times with PBS afterwards, nitrogen dries up, and obtains core (silver-colored)-satellite (golden) nano-particle and repaiies
The SERS chip of the silicon wafer of decorations.
Embodiment 3: prepare silica-based SERS chip of the present invention
(1) Fluohydric acid. auxiliary etch method prepares the silicon wafer of modified by silver nanoparticles
Take 0.5cm2Size monocrystalline silicon piece 3-6 sheet is placed in clean beaker and spends successively in Ultrasound Instrument
Ionized water, acetone, deionized water carry out ultrasonic cleaning 15 minutes, place into 40mL concentrated sulphuric acid and
Hydrogen peroxide (mass concentration: 40%) mixed solution (volume ratio=3:1) cleans further,
Clean with deionized water the most again, obtain clean silicon wafer.The silicon wafer cleaned up is put
Enter in hydrofluoric acid solution (mass concentration: 5%) and carry out silicon-hydrogenation, slowly vibration 30 minutes,
Obtain surface and cover the silicon wafer of a large amount of Si--H bond.By silicon wafer obtained after above-mentioned process
Sheet, light faces up, and puts into 20mL silver nitrate (1M) and Fluohydric acid. (mass concentration: 40%)
Mixed solution (volume ratio=1:100) in, slow oscillating reactions 60 minutes, anti-according to electrochemistry
Principle, silver ion is answered to be reduced by Si--H bond, uniform at silicon wafer surface growth in situ last layer
Silver nano-grain, thus obtain the silicon wafer of modified by silver nanoparticles, finally dry up table with nitrogen
Face.
(2) preparation of the silicon wafer that core (silver-colored)-satellite (golden) is nano-particle modified
Citric acid method of reducing according to standard, at the 50mL chlorauric acid solution seethed with excitement, (quality is dense
Degree: 0.01%) in, add 2mL sodium citrate solution (mass concentration: 1%), stirring reaction
After 15 minutes, obtain gold nano grain.By prepared gold nano grain be dissolved in phosphoric acid
Poly A30-P1DNA (5 ' in salt buffer (PBS)
-AAAAAAAAAAAAAAAAAAAAAAAAAAAAAATTTTTGATACAG
CTAATTCAGAATCATTTTGTGGA-3 ') (concentration: 0.01M), constant temperature 25 DEG C,
On constant temperature blending instrument, mixing is hatched 16 hours, then divides in solution and added one every 2 hours 3 times
Secondary 1M saline solution so that saline solution ultimate density is 0.1M, the most aging, obtains Poly
The gold nano grain that A30-P1 connects.By the silicon wafer of modified by silver nanoparticles that prepared with
The Poly A30-P2DNA (5 ' being dissolved in phosphate buffer
-AAAAAAAAAAAAAAAAAAAAAAAAAAAAAATTTTTTCCACAA
AATGATTCTGAATTAGCTGTATC-3 ') (concentration: 0.01M), constant temperature 25 DEG C,
On constant temperature blending instrument, mixing is hatched 16 hours, then divides in solution and added one every 2 hours 5 times
Secondary 1M saline solution so that saline solution ultimate density is 0.1M, the most aging, obtains Poly
The silicon wafer of the modified by silver nanoparticles that A30-P2 connects.Above-mentioned two step resulting materials are placed in miscellaneous
Handing in buffer, constant temperature 37 DEG C, on constant temperature blending instrument, mixing hatches 24 hours.Pass through DNA
Complementary pairing forms duplex structure, rinses several times with PBS, and then nitrogen dries up, and obtains core (silver-colored)
The silicon wafer that-satellite (golden) is nano-particle modified.
(3) the SERS chip structure of the silicon wafer that core (silver-colored)-satellite (golden) is nano-particle modified
Build
Silicon wafer nano-particle modified for core (silver-colored)-satellite (golden) is placed in centrifuge tube, adds
Enter enzyme strand DNA (Cy5-17E-SH, 5 '
-HS-(CH2)6-TTTCATCTCTTCTCCGAGCCGGTCGAAATAGTGAGT-C
Y5-3 ') solution of (concentration: 0.001M) so that solution submergence material.Centrifuge tube is put
In constant temperature blending instrument, 350 rpms, 25 DEG C of constant temperature, oscillating reactions 24 hours, make DNA
Terminal sulfhydryl group and gold, silver nanoparticle covalency form gold-sulfide linkage and silver-sulfide linkage, make DNA covalency even
Receive on the silicon wafer that core (silver-colored)-satellite (golden) is nano-particle modified.Then in solution, divide 5
Secondary added a 1M saline solution every 2 hours so that saline solution ultimate density is 0.1M, the most always
Change.By material take out, be dissolved in substrate strand DNA in hybridization buffer (17DS,
5 '-ACTCACTATrAGGAAGAGATG-3 ', rA (ribonucleotide adenosine)
Represent base A in ribonucleotide) (concentration: 0.001M), 37 DEG C of constant temperature, at constant temperature
In blending instrument, mixing is hatched 24 hours, forms DNA double chain structure by DNA complementary pairing, so
Rinsing several times with PBS afterwards, nitrogen dries up, and obtains core (silver-colored)-satellite (golden) nano-particle and repaiies
The SERS chip of the silicon wafer of decorations.
Embodiment 3: prepare silica-based SERS chip of the present invention
(1) Fluohydric acid. auxiliary etch method prepares the silicon wafer of modified by silver nanoparticles
Take 0.5cm2Size monocrystalline silicon piece 3-6 sheet is placed in clean beaker and spends successively in Ultrasound Instrument
Ionized water, acetone, deionized water carry out ultrasonic cleaning 15 minutes, place into 40mL concentrated sulphuric acid and
Hydrogen peroxide (mass concentration: 40%) mixed solution (volume ratio=3:1) cleans further,
Clean with deionized water the most again, obtain clean silicon wafer.The silicon wafer cleaned up is put
Enter in hydrofluoric acid solution (mass concentration: 5%) and carry out silicon-hydrogenation, slowly vibration 30 minutes,
Obtain surface and cover the silicon wafer of a large amount of Si--H bond.By silicon wafer obtained after above-mentioned process
Sheet, light faces up, and puts into 20mL silver nitrate (1M) and Fluohydric acid. (mass concentration: 40%)
Mixed solution (volume ratio=1:100) in, slow oscillating reactions 60 minutes, anti-according to electrochemistry
Principle, silver ion is answered to be reduced by Si--H bond, uniform at silicon wafer surface growth in situ last layer
Silver nano-grain, thus obtain the silicon wafer of modified by silver nanoparticles, finally dry up table with nitrogen
Face.
(2) preparation of the silicon wafer that core (silver-colored)-satellite (golden) is nano-particle modified
Citric acid method of reducing according to standard, at the 50mL chlorauric acid solution seethed with excitement, (quality is dense
Degree: 0.01%) in, add 2mL sodium citrate solution (mass concentration: 1%), stirring reaction
After 15 minutes, obtain gold nano grain.By prepared gold nano grain be dissolved in phosphoric acid
Poly A30-P1DNA (5 ' in salt buffer (PBS)
-AAAAAAAAAAAAAAAAAAAAAAAAAAAAAATTTTTGATACAG
CTAATTCAGAATCATTTTGTGGA-3 ') (concentration: 0.001M), constant temperature 25 DEG C,
On constant temperature blending instrument, mixing is hatched 16 hours, then divides in solution and added one every 2 hours 3 times
Secondary 1M saline solution so that saline solution ultimate density is 0.1M, the most aging, obtains Poly
The gold nano grain that A30-P1 connects.By the silicon wafer of modified by silver nanoparticles that prepared with
The Poly A30-P2DNA (5 ' being dissolved in phosphate buffer
-AAAAAAAAAAAAAAAAAAAAAAAAAAAAAATTTTTTCCACAA
AATGATTCTGAATTAGCTGTATC-3 ') (concentration: 0.001M), constant temperature 25 DEG C,
On constant temperature blending instrument, mixing is hatched 16 hours, then divides in solution and added one every 2 hours 5 times
Secondary 1M saline solution so that saline solution ultimate density is 0.1M, the most aging, obtains Poly
The silicon wafer of the modified by silver nanoparticles that A30-P2 connects.Above-mentioned two step resulting materials are placed in miscellaneous
Handing in buffer, constant temperature 37 DEG C, on constant temperature blending instrument, mixing hatches 24 hours.Pass through DNA
Complementary pairing forms duplex structure, rinses several times with PBS, and then nitrogen dries up, and obtains core (silver-colored)
The silicon wafer that-satellite (golden) is nano-particle modified.
(3) the SERS chip structure of the silicon wafer that core (silver-colored)-satellite (golden) is nano-particle modified
Build
Silicon wafer nano-particle modified for core (silver-colored)-satellite (golden) is placed in centrifuge tube, adds
Enter enzyme strand DNA (Cy5-17E-SH, 5 '
-HS-(CH2)6-TTTCATCTCTTCTCCGAGCCGGTCGAAATAGTGAGT-C
Y5-3 ') solution of (concentration: 0.01M) so that solution submergence material.Centrifuge tube is put
In constant temperature blending instrument, 350 rpms, 25 DEG C of constant temperature, oscillating reactions 24 hours, make DNA
Terminal sulfhydryl group and gold, silver nanoparticle covalency form gold-sulfide linkage and silver-sulfide linkage, make DNA covalency even
Receive on the silicon wafer that core (silver-colored)-satellite (golden) is nano-particle modified.Then in solution, divide 5
Secondary added a 1M saline solution every 2 hours so that saline solution ultimate density is 0.1M, the most always
Change.By material take out, be dissolved in substrate strand DNA in hybridization buffer (17DS,
5 '-ACTCACTATrAGGAAGAGATG-3 ', rA (ribonucleotide adenosine)
Represent base A in ribonucleotide) (concentration: 0.01M), 37 DEG C of constant temperature, at constant temperature
In blending instrument, mixing is hatched 24 hours, forms DNA double chain structure by DNA complementary pairing, so
Rinsing several times with PBS afterwards, nitrogen dries up, and obtains core (silver-colored)-satellite (golden) nano-particle and repaiies
The SERS chip of the silicon wafer of decorations.
Embodiment 5: prepare silica-based SERS chip of the present invention
(1) Fluohydric acid. auxiliary etch method prepares the silicon wafer of modified by silver nanoparticles
Take 0.5cm2Size monocrystalline silicon piece 3-6 sheet is placed in clean beaker and spends successively in Ultrasound Instrument
Ionized water, acetone, deionized water carry out ultrasonic cleaning 15 minutes, place into 40mL concentrated sulphuric acid and
Hydrogen peroxide (mass concentration: 40%) mixed solution (volume ratio=3:1) cleans further,
Clean with deionized water the most again, obtain clean silicon wafer.The silicon wafer cleaned up is put
Enter in hydrofluoric acid solution (mass concentration: 5%) and carry out silicon-hydrogenation, slowly vibration 30 minutes,
Obtain surface and cover the silicon wafer of a large amount of Si--H bond.By silicon wafer obtained after above-mentioned process
Sheet, light faces up, and puts into 20mL silver nitrate (1M) and Fluohydric acid. (mass concentration: 40%)
Mixed solution (volume ratio=1:100) in, slow oscillating reactions 60 minutes, anti-according to electrochemistry
Principle, silver ion is answered to be reduced by Si--H bond, uniform at silicon wafer surface growth in situ last layer
Silver nano-grain, thus obtain the silicon wafer of modified by silver nanoparticles, finally dry up table with nitrogen
Face.
(2) preparation of the silicon wafer that core (silver-colored)-satellite (golden) is nano-particle modified
Citric acid method of reducing according to standard, at the 50mL chlorauric acid solution seethed with excitement, (quality is dense
Degree: 0.01%) in, add 2mL sodium citrate solution (mass concentration: 1%), stirring reaction
After 15 minutes, obtain gold nano grain.By prepared gold nano grain be dissolved in phosphoric acid
Poly A30-P1DNA (5 ' in salt buffer (PBS)
-AAAAAAAAAAAAAAAAAAAAAAAAAAAAAATTTTTGATACAG
CTAATTCAGAATCATTTTGTGGA-3 ') (concentration: 0.001M), constant temperature 25 DEG C,
On constant temperature blending instrument, mixing is hatched 16 hours, then divides in solution and added one every 2 hours 3 times
Secondary 1M saline solution so that saline solution ultimate density is 0.5M, the most aging, obtains Poly
The gold nano grain that A30-P1 connects.By the silicon wafer of modified by silver nanoparticles that prepared with
The Poly A30-P2DNA (5 ' being dissolved in phosphate buffer
-AAAAAAAAAAAAAAAAAAAAAAAAAAAAAATTTTTTCCACAA
AATGATTCTGAATTAGCTGTATC-3 ') (concentration: 0.001M), constant temperature 25 DEG C,
On constant temperature blending instrument, mixing is hatched 16 hours, then divides in solution and added one every 2 hours 5 times
Secondary 1M saline solution so that saline solution ultimate density is 0.5M, the most aging, obtains Poly
The silicon wafer of the modified by silver nanoparticles that A30-P2 connects.Above-mentioned two step resulting materials are placed in miscellaneous
Handing in buffer, constant temperature 37 DEG C, on constant temperature blending instrument, mixing hatches 24 hours.Pass through DNA
Complementary pairing forms duplex structure, rinses several times with PBS, and then nitrogen dries up, and obtains core (silver-colored)
The silicon wafer that-satellite (golden) is nano-particle modified.
(3) the SERS chip structure of the silicon wafer that core (silver-colored)-satellite (golden) is nano-particle modified
Build
Silicon wafer nano-particle modified for core (silver-colored)-satellite (golden) is placed in centrifuge tube, adds
Enter enzyme strand DNA (Cy5-17E-SH, 5 '
-HS-(CH2)6-TTTCATCTCTTCTCCGAGCCGGTCGAAATAGTGAGT-C
Y5-3 ') solution of (concentration: 0.001M) so that solution submergence material.Centrifuge tube is put
In constant temperature blending instrument, 350 rpms, 25 DEG C of constant temperature, oscillating reactions 24 hours, make DNA
Terminal sulfhydryl group and gold, silver nanoparticle covalency form gold-sulfide linkage and silver-sulfide linkage, make DNA covalency even
Receive on the silicon wafer that core (silver-colored)-satellite (golden) is nano-particle modified.Then in solution, divide 5
Secondary added a 1M saline solution every 2 hours so that saline solution ultimate density is 0.5M, the most always
Change.By material take out, be dissolved in substrate strand DNA in hybridization buffer (17DS,
5 '-ACTCACTATrAGGAAGAGATG-3 ', rA (ribonucleotide adenosine)
Represent base A in ribonucleotide) (concentration: 0.001M), 37 DEG C of constant temperature, at constant temperature
In blending instrument, mixing is hatched 24 hours, forms DNA double chain structure by DNA complementary pairing, so
Rinsing several times with PBS afterwards, nitrogen dries up, and obtains core (silver-colored)-satellite (golden) nano-particle and repaiies
The SERS chip of the silicon wafer of decorations.
Embodiment 6: prepare silica-based SERS chip of the present invention
(1) Fluohydric acid. auxiliary etch method prepares the silicon wafer of modified by silver nanoparticles
Take 0.5cm2Size monocrystalline silicon piece 3-6 sheet is placed in clean beaker and spends successively in Ultrasound Instrument
Ionized water, acetone, deionized water carry out ultrasonic cleaning 15 minutes, place into 40mL concentrated sulphuric acid and
Hydrogen peroxide (mass concentration: 40%) mixed solution (volume ratio=3:1) cleans further,
Clean with deionized water the most again, obtain clean silicon wafer.The silicon wafer cleaned up is put
Enter in hydrofluoric acid solution (mass concentration: 5%) and carry out silicon-hydrogenation, slowly vibration 30 minutes,
Obtain surface and cover the silicon wafer of a large amount of Si--H bond.By silicon wafer obtained after above-mentioned process
Sheet, light faces up, and puts into 20mL silver nitrate (1M) and Fluohydric acid. (mass concentration: 40%)
Mixed solution (volume ratio=1:100) in, slow oscillating reactions 60 minutes, anti-according to electrochemistry
Principle, silver ion is answered to be reduced by Si--H bond, uniform at silicon wafer surface growth in situ last layer
Silver nano-grain, thus obtain the silicon wafer of modified by silver nanoparticles, finally dry up table with nitrogen
Face.
(2) preparation of the silicon wafer that core (silver-colored)-satellite (golden) is nano-particle modified
Citric acid method of reducing according to standard, at the 50mL chlorauric acid solution seethed with excitement, (quality is dense
Degree: 0.01%) in, add 2mL sodium citrate solution (mass concentration: 1%), stirring reaction
After 15 minutes, obtain gold nano grain.By prepared gold nano grain be dissolved in phosphoric acid
Poly A30-P1DNA (5 ' in salt buffer (PBS)
-AAAAAAAAAAAAAAAAAAAAAAAAAAAAAATTTTTGATACAG
CTAATTCAGAATCATTTTGTGGA-3 ') (concentration: 0.001M), constant temperature 25 DEG C,
On constant temperature blending instrument, mixing is hatched 16 hours, then divides in solution and added one every 2 hours 3 times
Secondary 1M saline solution so that saline solution ultimate density is 0.1M, the most aging, obtains Poly
The gold nano grain that A30-P1 connects.By the silicon wafer of modified by silver nanoparticles that prepared with
The Poly A30-P2DNA (5 ' being dissolved in phosphate buffer
-AAAAAAAAAAAAAAAAAAAAAAAAAAAAAATTTTTTCCACAA
AATGATTCTGAATTAGCTGTATC-3 ') (concentration: 0.001M), constant temperature 25 DEG C,
On constant temperature blending instrument, mixing is hatched 16 hours, then divides in solution and added one every 2 hours 5 times
Secondary 1M saline solution so that saline solution ultimate density is 0.1M, the most aging, obtains Poly
The silicon wafer of the modified by silver nanoparticles that A30-P2 connects.Above-mentioned two step resulting materials are placed in miscellaneous
Handing in buffer, constant temperature 37 DEG C, on constant temperature blending instrument, mixing hatches 24 hours.Pass through DNA
Complementary pairing forms duplex structure, rinses several times with PBS, and then nitrogen dries up, and obtains core (silver-colored)
The silicon wafer that-satellite (golden) is nano-particle modified.
(3) the SERS chip structure of the silicon wafer that core (silver-colored)-satellite (golden) is nano-particle modified
Build
Silicon wafer nano-particle modified for core (silver-colored)-satellite (golden) is placed in centrifuge tube, adds
Enter enzyme strand DNA (Cy5-17E-SH, 5 '
-HS-(CH2)6-TTTCATCTCTTCTCCGAGCCGGTCGAAATAGTGAGT-Cy
5-3 ') solution of (concentration: 0.001M) so that solution submergence material.Centrifuge tube is placed in
In constant temperature blending instrument, 500 rpms, 25 DEG C of constant temperature, oscillating reactions 18 hours, make DNA
Terminal sulfhydryl group and gold, silver nanoparticle covalency form gold-sulfide linkage and silver-sulfide linkage, make DNA covalency
It is connected on the silicon wafer that core (silver-colored)-satellite (golden) is nano-particle modified.Then in solution
Divide 5 times and added a 1M saline solution every 2 hours so that saline solution ultimate density is 0.1M,
The most aging.Material is taken out, and is dissolved in substrate strand DNA in hybridization buffer
(17DS, 5 '-ACTCACTATrAGGAAGAGATG-3 ', rA (ribonucleotide
Adenosine) base A in ribonucleotide is represented) (concentration: 0.001M), 37 DEG C of perseverances
Temperature, mixes in constant temperature blending instrument and hatches 18 hours, forms DNA by DNA complementary pairing
Duplex structure, then rinses several times with PBS, and nitrogen dries up, and obtains core (silver-colored)-satellite (golden)
The SERS chip of nano-particle modified silicon wafer.
Embodiment 7: the SERS of variable concentrations lead ion is tested by chip of the present invention
By the chip of the present invention for preparing at room temperature, it is immersed in 100nM lead ion solution,
After reacting 70 minutes, take out chip, randomly choose 40 points and do mapping experiment, by Fig. 3
It is more uniform that middle a figure understands raman scattering intensity, and RSD value is less than 12%, indicates this chip and has
Preferably repeatability.
By the chip of the present invention for preparing at room temperature, be separately immersed in 0,10pM, 100pM,
In 1nM, 10nM, 100nM, 1 μM of lead ion solution, after reacting 70 minutes, take out core
Sheet, carries out Raman experiments, b figure in Fig. 3 understand the Raman from 10pM to 1 μM of lead ion
Spectrogram, is also increasing along with plumbum ion concentration increases raman scattering intensity, c in Fig. 3 is scheming
There is preferable linear relationship in the log concentration of lead ion and normalized raman scattering intensity
(R2=0.997), test result indicate that this chip can effectively detect plumbum ion concentration as little as 8.9pM,
Sensitivity is high.
Embodiment 8: the SERS of same concentration different ions is tested by chip of the present invention
The chip prepared is separately immersed at room temperature the various ions of 1nM and various
In the mixed solution of ion, after reacting 70 minutes, take out chip, carry out Raman experiments respectively,
Schemed to understand lead ion by a in Fig. 4 to compare other ions with containing the mixed solution of lead ion
The Raman spectrum of solution has significantly enhancing, and the b figure from Fig. 4 can be seen that and draws accordingly
Graceful peak 1366cm-1Intensity can be quantitative comparison different ions between difference, result shows this
Chip has preferable specificity can accurately identify the lead ion in practical systems.
Embodiment 9: the reconstruction of chip of the present invention and recycling test
1, the reconstruction of chip of the present invention
After one-time detection, with PBS chip to remove remaining DNA and lead ion,
By material take out, be dissolved in substrate strand DNA in hybridization buffer (17DS, 5 '
-ACTCACTATrAGGAAGAGATG-3 ', rA (ribonucleotide adenosine)
Represent base A in ribonucleotide) (concentration: 0.001M), 37 DEG C of constant temperature, at constant temperature
In blending instrument, mixing is hatched 24 hours, forms DNA double chain structure by DNA complementary pairing,
Then rinsing several times with PBS, nitrogen dries up, and rebuilds and obtains core (silver-colored)-satellite (golden) nanometer
The SERS chip (seeing a figure in Fig. 5) of the silicon wafer of particle modification.
2, the recycling test of chip of the present invention
The chip that will rebuild, at room temperature, is immersed in the lead ion solution of 1nM, reaction
After 70 minutes, take out chip, carry out Raman experiments, then recycle, rebuild it each time
Rear raman scattering intensity all can significantly strengthen that (rebuild every time after, raman scattering intensity all can be remarkably reinforced finger
It is after just rebuilding there there is no or the faintest, when adding lead ion the raman scattering intensity of untapped chip
Raman signal can significantly strengthen afterwards, sees b figure in Fig. 5), for quantitative comparison, by scheming
Raman peaks 1366cm in c figure in 5-1Strength co-mputation learn three times circulation after intensity only weaken
11%, result shows that chip of the present invention is prone to rebuild, and can be used for multiple times, and recyclability is high,
It is substantially reduced preparation cost.
The above is only the preferred embodiment of the present invention, it is noted that lead for this technology
For the those of ordinary skill in territory, under the premise without departing from the principles of the invention, it is also possible to make
Some improvements and modifications, these improvements and modifications also should be regarded as protection scope of the present invention.
Claims (18)
1. the silica-based SERS chip of plumbum ion concentration in a detection by quantitative actual water sample, it is characterised in that
By the silicon wafer of modified by silver nanoparticles, gold nano grain, SEQ ID NO:1-2 and SEQ ID NO:4
Shown in nucleotide sequence, and nucleotide sequence 5 ' shown in SEQ ID NO:3 end coupling have sulfydryl, 3 '
End coupling is made up of the sequence fragment of fluorescent dye；
Wherein, the silicon wafer of modified by silver nanoparticles is connected with nucleotide sequence shown in SEQ ID NO:1,
Gold nano grain is connected with nucleotide sequence shown in SEQ ID NO:2, nucleoside shown in SEQ ID NO:1-2
Acid sequence forms complementary double-strand each other and connects, and has in the end coupling of nucleotide sequence 5 ' shown in SEQ ID NO:3
Sulfydryl, 3 ' end couplings have the sequence fragment of fluorescent dye by terminal sulfhydryl group and gold, silver nano-particle covalency
Connect, nucleotide sequence shown in SEQ ID NO:4 and nucleotide sequence shape each other shown in SEQ ID NO:3
Complementary double-strand is become to connect.
The most silica-based SERS chip, it is characterised in that described sequence fragment is
HS-(CH is had in the end coupling of nucleotide sequence 5 ' shown in SEQ ID NO:32)6-, 3 ' end couplings have Cy5 glimmering
The sequence fragment of photoinitiator dye.
The most silica-based SERS chip, it is characterised in that described silicon wafer is
The p-type of 0.01～20 Ω * cm or n-type silicon wafer.
4. the preparation method of silica-based SERS chip described in claim 1, it is characterised in that including:
Step 1, the silicon wafer preparing modified by silver nanoparticles and gold nano grain；
Step 2, by the silicon wafer of the modified by silver nanoparticles prepared be dissolved in phosphate buffer
In SEQ ID NO:1 shown in nucleotide sequence constant temperature mixing hatch, be then added thereto to saline solution,
The most aging, obtain the silicon wafer of the modified by silver nanoparticles that nucleotide sequence shown in SEQ ID NO:1 connects
Sheet；
By prepared gold nano grain and core shown in the SEQ ID NO:2 being dissolved in phosphate buffer
The mixing of nucleotide sequence constant temperature is hatched, and then adds saline solution in solution, the most aging, obtains SEQ ID
The gold nano grain that nucleotide sequence shown in NO:2 connects；
Step 3, step 2 gained silicon wafer and gold nano grain are placed in hybridization buffer constant temperature mixing
Hatch, form duplex structure by the pairing of nucleotide sequence complementary shown in SEQ ID NO:1-2, use PBS
Rinsing, then nitrogen dries up, and obtains the silicon wafer that core (silver-colored)-satellite (golden) is nano-particle modified；
Step 4, by silicon wafer nano-particle modified for core (silver-colored)-satellite (golden) with at SEQ ID NO:3
Shown nucleotide sequence 5 ' end coupling has sulfydryl, 3 ' end couplings to have the sequence fragment constant temperature blending of fluorescent dye
And oscillating reactions, make nucleotide sequence terminal sulfhydryl group shown in SEQ ID NO:3 with gold, silver nanoparticle altogether
Valency connects formation gold-sulfide linkage and silver-sulfide linkage, then adds saline solution in solution, the most aging；
Material after aging is taken out, and is dissolved in nucleotide shown in SEQ ID NO:4 in hybridization buffer
The mixing of sequence constant temperature is hatched, and forms DNA double chain structure by DNA complementary pairing, then uses PBS
Rinsing, nitrogen dries up, and obtains described silica-based SERS chip.
Preparation method the most according to claim 4, it is characterised in that described modified by silver nanoparticles
Silicon wafer is prepared by following methods:
Monocrystalline silicon piece is carried out ultrasonic cleaning with deionized water, acetone, deionized water successively, uses the most again
Concentrated sulphuric acid and mixed solution of hydrogen peroxide clean；
Monocrystalline silicon piece after cleaning joins and carries out silicon-hydrogenation in hydrofluoric acid solution, obtains surface and covers
The silicon wafer of Si--H bond, then light faces up, and puts in the mixed solution of silver nitrate and Fluohydric acid., slowly
Oscillating reactions, silver ion is reduced by Si--H bond, receives at silicon wafer surface growth in situ last layer silver uniformly
Rice grain, obtains the silicon wafer of modified by silver nanoparticles, finally dries up surface with nitrogen.
Preparation method the most according to claim 5, it is characterised in that described hydrogen peroxide quality is dense
Degree is 40%, and concentrated sulphuric acid and 40% hydrogen peroxide volume ratio are 1:(0.01～100).
Preparation method the most according to claim 5, it is characterised in that hydrogen fluorine in described hydrofluoric acid solution
The mass concentration of acid is 1～40%.
Preparation method the most according to claim 5, it is characterised in that described silicon-hydrogenation time
Between be 1～60 minute.
Preparation method the most according to claim 5, it is characterised in that described silver nitrate and Fluohydric acid.
Mixed solution is 1 by silver nitrate solution and the hydrofluoric acid solution that mass concentration is 40% of 1M by volume:
(0.01～100) are formulated.
Preparation method the most according to claim 5, it is characterised in that the described oscillating reactions time is
1～60 minute.
11. preparation methoies according to claim 4, it is characterised in that described nanogold particle passes through
Citric acid reducing process prepares.
12. preparation methoies according to claim 4, it is characterised in that shown in SEQ ID NO:1-2
Nucleotide sequence concentration is 0.001～1M.
13. preparation methoies according to claim 4, it is characterised in that constant temperature mixing described in step 2
Hatch as hatching 16 hours at 25 DEG C.
14. preparation methoies according to claim 4, it is characterised in that described in step 2 and step 4
Adding saline solution is to be the saline solution of 1M by initial concentration, and every 3-5 the addition of two little time-divisions, salt is molten
Liquid ultimate density is 0.01～1M.
15. preparation methoies according to claim 4, it is characterised in that constant temperature mixing described in step 3
Hatch as hatching 24 hours at 37 DEG C.
16. preparation methoies according to claim 4, it is characterised in that oscillating reactions described in step 4
For 100～600 rpms, react 1～24 hour at 25 DEG C.
17. preparation methoies according to claim 4, it is characterised in that constant temperature mixing described in step 4
Hatch as hatching 1～24 hour at 37 DEG C.
18. preparation methoies according to claim 4, it is characterised in that at SEQ ID NO:3 in step 4
Shown nucleotide sequence 5 ' end coupling has sulfydryl, 3 ' end couplings to have the sequence fragment of fluorescent dye, SEQ ID
Nucleotide sequence concentration shown in NO:4 is 0.001～1M.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106323934A (en) * | 2016-11-18 | 2017-01-11 | 重庆工商大学 | Fluorescent biological probe for simultaneous determination of three ions of Cu2 +, Mg2 + and Pb2 + and detecting method thereof |
| CN106645364A (en) * | 2017-02-27 | 2017-05-10 | 福州大学 | Method for detecting Cu2+ on basis of polyadenine-methylene blue electrochemical sensor |
| CN109187992A (en) * | 2018-09-12 | 2019-01-11 | 天津科技大学 | A kind of label-free fluorescent optical sensor of novel Ratio-type and its application |
| CN110106226A (en) * | 2019-06-04 | 2019-08-09 | 中国工程物理研究院化工材料研究所 | Bio-sensing chip of recyclable detection trace uranyl ion and preparation method thereof, application method |
| CN111534571A (en) * | 2020-05-25 | 2020-08-14 | 长江师范学院 | CHA-SERS biosensor for lead ion detection and preparation method and application thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1553401A1 (en) * | 2004-01-07 | 2005-07-13 | Fuji Photo Film Co., Ltd | Raman spectroscopy method and device |
| US20140002816A1 (en) * | 2012-06-29 | 2014-01-02 | National Institute For Materials Science | Substrate for surface enhanced raman spectroscopy analysis and manufacturing method of the same, biosensor using the same, and microfluidic device using the same |
| CN103604796A (en) * | 2013-11-29 | 2014-02-26 | 苏州大学 | Preparation method for silicon-based surface-enhanced Raman scattering (SERS) substrate |
| CN104215626A (en) * | 2014-09-24 | 2014-12-17 | 苏州大学 | Method for detecting deafness genes based on surface enhanced Raman spectroscopy |
| CN104237203A (en) * | 2014-09-28 | 2014-12-24 | 苏州大学 | SERS sensor for quantitatively detecting concentration of mercury ions in water sample and preparation method of SERS sensor |
| CN104697977A (en) * | 2015-03-23 | 2015-06-10 | 苏州大学 | Silicon-based SERS multifunctional chip and preparation method thereof |
-
2016
- 2016-03-28 CN CN201610179491.XA patent/CN105842225A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1553401A1 (en) * | 2004-01-07 | 2005-07-13 | Fuji Photo Film Co., Ltd | Raman spectroscopy method and device |
| US20140002816A1 (en) * | 2012-06-29 | 2014-01-02 | National Institute For Materials Science | Substrate for surface enhanced raman spectroscopy analysis and manufacturing method of the same, biosensor using the same, and microfluidic device using the same |
| CN103604796A (en) * | 2013-11-29 | 2014-02-26 | 苏州大学 | Preparation method for silicon-based surface-enhanced Raman scattering (SERS) substrate |
| CN104215626A (en) * | 2014-09-24 | 2014-12-17 | 苏州大学 | Method for detecting deafness genes based on surface enhanced Raman spectroscopy |
| CN104237203A (en) * | 2014-09-28 | 2014-12-24 | 苏州大学 | SERS sensor for quantitatively detecting concentration of mercury ions in water sample and preparation method of SERS sensor |
| CN104697977A (en) * | 2015-03-23 | 2015-06-10 | 苏州大学 | Silicon-based SERS multifunctional chip and preparation method thereof |
Non-Patent Citations (2)
| Title |
|---|
| YU SHI ET AL.: "Ultrasensitive, Specific, Recyclable, and Reproducible Detection of Lead Ions in Real Systems through a Polyadenine-Assisted, Surface-Enhanced Raman Scattering Silicon Chip", 《ANALYTICAL CHEMISTRY》 * |
| 朱瀛: "基于多聚腺嘌呤技术构建新型高效硅基SERS传感器及其检测实体水中汞离子的应用", 《中国优秀硕士学位论文全文数据库 工程科技I辑》 * |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106323934A (en) * | 2016-11-18 | 2017-01-11 | 重庆工商大学 | Fluorescent biological probe for simultaneous determination of three ions of Cu2 +, Mg2 + and Pb2 + and detecting method thereof |
| CN106323934B (en) * | 2016-11-18 | 2019-06-14 | 重庆工商大学 | It is a kind of to detect Cu simultaneously2+、Mg2+And Pb2+The fluorescent bio-probes and its detection method of three kinds of ions |
| CN106645364A (en) * | 2017-02-27 | 2017-05-10 | 福州大学 | Method for detecting Cu2+ on basis of polyadenine-methylene blue electrochemical sensor |
| CN109187992A (en) * | 2018-09-12 | 2019-01-11 | 天津科技大学 | A kind of label-free fluorescent optical sensor of novel Ratio-type and its application |
| CN109187992B (en) * | 2018-09-12 | 2021-09-07 | 天津科技大学 | Method for detecting lead ions by ratio type label-free fluorescence |
| CN110106226A (en) * | 2019-06-04 | 2019-08-09 | 中国工程物理研究院化工材料研究所 | Bio-sensing chip of recyclable detection trace uranyl ion and preparation method thereof, application method |
| CN111534571A (en) * | 2020-05-25 | 2020-08-14 | 长江师范学院 | CHA-SERS biosensor for lead ion detection and preparation method and application thereof |
| CN111534571B (en) * | 2020-05-25 | 2022-10-11 | 长江师范学院 | CHA-SERS biosensor for lead ion detection and preparation method and application thereof |
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