CN106086030A - Sandwich structure Raman signal for fingerprint imaging strengthens probe and preparation method thereof - Google Patents
Sandwich structure Raman signal for fingerprint imaging strengthens probe and preparation method thereof Download PDFInfo
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- CN106086030A CN106086030A CN201610396569.3A CN201610396569A CN106086030A CN 106086030 A CN106086030 A CN 106086030A CN 201610396569 A CN201610396569 A CN 201610396569A CN 106086030 A CN106086030 A CN 106086030A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/115—Aptamers, i.e. nucleic acids binding a target molecule specifically and with high affinity without hybridising therewith ; Nucleic acids binding to non-nucleic acids, e.g. aptamers
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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
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/10—Type of nucleic acid
- C12N2310/16—Aptamers
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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
- G01N2021/653—Coherent methods [CARS]
- G01N2021/656—Raman microprobe
Abstract
The present invention relates to a kind of sandwich structure Raman signal for fingerprint imaging and strengthen probe and preparation method thereof, this probe is with gold nano grain as core, silicon dioxide is shell, and signaling molecule is sandwich between described core and described shell, and described shell surface is modified with the lysozyme aptamers for fingerprint recognition.Its preparation method includes step: (1) prepares gold nano grain；(2) by S Au key, signaling molecule is connected on described gold nano grain；(3) silicon layer is superscribed by sodium silicate hydrolysis；(4) by forming triazine radical by amido modified lysozyme aptamers function on its surface in silicon surface.The sandwich structure probe of the present invention has that the highest Raman signal reinforced effects, high signal stabilization, high fingerprint recognition feature, and described preparation method be simple, process control, and prepared Raman signal strengthens probe and can be used for diving the high definition imaging of fingerprint.
Description
Technical field
The invention belongs to technical field of function materials, be specifically related to a kind of sandwich structure Raman signal for fingerprint imaging
Strengthen probe and preparation method thereof.
Background technology
Fingerprint is the skin texture that human finger's end refers on abdomen, by ' ridge line ' and ' the valley line ' of depression of a series of projections
Spaced apart form.Fingerprint varies with each individual, different because of finger, there is uniqueness；And fingerprint is once formed, its feature is lifelong not
Become, there is relative stability.Therefore fingerprint identification has been widely used for criminal detection and person identification.Fingerprint identification is
Carried out by relatively different fingerprint details.In general the feature of fingerprint is divided into Three Estate, i.e. pattern
(pattern), details (minutia points) and fine-feature (pores and ridge contours).And details and essence
Fine texture has uniqueness, is the foundation of personal identification qualification.But the visualization of details and fine structure is challenging, especially
It is the visualization of fine structure, needs by means of the imaging technique with high-resolution.Further, present most of fingerprints become
As technology has selectivity to detection substrate, versatility is poor.Therefore, a kind of high-resolution, fingerprint imaging of generalization is developed
Strategy causes in chemistry and forensic science and studies interest widely.
Surface Enhanced Raman Scattering Spectrum (SERS) is that a kind of hypersensitive grown up in the technology of Raman spectrum shakes
Dynamic spectrum, which overcome Raman spectrum intrinsic cannot detect low concentration material and the shortcoming of background fluorescence interference, there is many
Significantly advantage: (1) can provide the chemical constitution on molecular level and functional group's information；(2) breadth of spectrum line is narrow, is suitable for polynary point
Analysis；(3) do not disturbed by hydrone, can be used for aqueous sample detection；(4) detection sensitivity is high, it might even be possible to reach unimolecule
Level.And SERS imaging technique be a kind of lossless, without photobleaching, high-resolution imaging technology, be widely used in biology
Material detection and visualization (Analytical Chemistry, the 2014,86,11503-11507 of metabolic pathway；
Analytical Chemistry,2014,86,4775-4782；Nature Protocols,2013,8,677-692).
Aptamers identification technology is a kind of high sensitivity, high Selective recognition technology, compared with antigen-antibody identification technology,
The identification of aptamers has the motility of some obvious advantages, such as design route, the tolerance of synthesis condition, easily modification
And biochemical stability.Lysozyme is a kind of polypeptide being present in human sweat, the skin surfaces such as mankind's forehead or the wing of nose with
And fingerprint ridge line the most generally exists.Its aptamers, as the identification body of fingerprint imaging, can realize the high-resolution imaging of fingerprint.
In conjunction with SERS imaging technique and the advantage of aptamers identification technology, exploitation preparation is applicable to fingerprint high definition imaging, and
And the Raman signal enhancing probe of preparation operation easy, easy, signal stabilization becomes the focus of this area research in recent years, but use
Present in latent fingerprint, lysozyme carries out SERS imaging identification as target molecule, up to now, there is not yet relevant report.
Summary of the invention
One aspect of the present invention provides a kind of sandwich structure Raman signal for fingerprint imaging and strengthens probe.The present invention's
Sandwich structure probe for fingerprint high definition imaging is with gold nano grain as core, and silicon dioxide is shell, signaling molecule sandwich in
Between described core and described shell, described shell surface is modified with the lysozyme aptamers for fingerprint recognition.
The sandwich structure probe for fingerprint high definition imaging of the present invention has the raman spectral signal of enhancing, therein molten
Bacterium enzyme aptamers is short single stranded oligonucleotide, and when not having lysozyme to occur, it can remain the state of strand；But when molten
When bacterium enzyme occurs, it can be folded into special complicated three dimensional structure, is then bonded in the surface of lysozyme, thus by sandwich knot
Structure SERS deposited probes is on the surface of fingerprint, it is achieved the detection of fingerprint.
In another preference, described signaling molecule is p-Nitrobenzenethiol, mercaptopyridine or mercaptobenzoic acid.
In another preference, the sequence of described lysozyme aptamers is 5 '-NH2-TTT TTT ATC AGG GCT AAA
GAG TGC AGA GTT ACT TAG-3'.It should be understood that the method that the present invention provides has universality, it is only necessary to according to this
First bright spirit select aptamers corresponding with lysozyme.More preferably.Aptamers is the 5 ' ends at Single-stranded DNA fragments
Add some T bases respectively, such as 5-10.Meanwhile, 5 ' ends of this Single-stranded DNA fragments are amido modified.Sequence is 5 '-NH2-
The lysozyme aptamers of TTT TTT ATC AGG GCT AAA GAG TGC AGA GTT ACT TAG-3' is only used as preferred reality
Execute and exemplify.
In another preference, a diameter of the 52.6 of described gold nano grain～58.6nm.Owing to nanometer gold itself has
Good biocompatibility so that the sandwich structure probe for fingerprint high definition imaging provided by the present invention has good life
The thing compatibility, so that probe provided by the present invention does not has any toxic action to operator.Preparing of nanometer gold is permissible
Carry out according to known method of the prior art.It practice, this nanometer gold can be the gold nano of any size, shape
Grain, above-mentioned a diameter of 52.6～58.6nm gold nano grain be only used as being preferable to carry out exemplifying.
In another preference, the thickness of silicon layer is 2.5～10nm.The thickness effect laser of silicon layer is delivered to gold from shell
The efficiency of core, the most ultra-thin silicon layer is that sandwich structure Raman signal strengthens probe for fingerprint visualization successfully key.On
Stating thickness is that 2.5～10nm silicon layers are only used as being preferable to carry out exemplifying.
In another preference, the lysozyme aptamers that single sandwich structure Raman signal enhancing detecting probe surface is modified
Number is 317～351.The number of the lysozyme aptamers that detecting probe surface is modified directly affects fingerprint detection efficiency.Due to letter
Number molecule is sandwich in silicon layer, and entirely free on his silicon surface can modify the maximum amount of lysozyme aptamers, significantly improves
Recognition efficiency.The number of single detecting probe surface lysozyme aptamers is can be evaluated whether according to Lambert-Beer's law.Above-mentioned single probe
The number of surface lysozyme aptamers is 317～351 to be only used as being preferable to carry out exemplifying.
Another aspect of the present invention provides a kind of sandwich structure Raman signal for fingerprint imaging and strengthens the preparation of probe
Method, the method comprises the steps:
(1) gold nano grain is prepared；(2) by S-Au key, signaling molecule is connected on described gold nano grain；(3) logical
Cross sodium silicate hydrolysis and superscribe silicon layer；(4) by forming triazine radical by amido modified lysozyme aptamers merit in silicon surface
Can be on its surface.
In another preference, described signaling molecule is p-Nitrobenzenethiol, mercaptopyridine or mercaptobenzoic acid.
In another preference, the sequence of described lysozyme aptamers is 5 '-NH2-TTT TTT ATC AGG GCT AAA
GAG TGC AGA GTT ACT TAG-3'.It should be understood that the method that the present invention provides has universality, it is only necessary to according to this
First bright spirit select aptamers corresponding with lysozyme.More preferably.Aptamers is the 5 ' ends at Single-stranded DNA fragments
Add some T bases respectively, such as 5-10.Meanwhile, 5 ' ends of this Single-stranded DNA fragments are amido modified.Sequence is 5 '-NH2-
The lysozyme aptamers of TTT TTT ATC AGG GCT AAA GAG TGC AGA GTT ACT TAG-3' is only used as preferred reality
Execute and exemplify.
In another preference, a diameter of the 52.6 of described gold nano grain～58.6nm.Owing to nanometer gold itself has
Good biocompatibility so that the sandwich structure probe for fingerprint high definition imaging provided by the present invention has good life
The thing compatibility, so that probe provided by the present invention does not has any toxic action to operator.Preparing of nanometer gold is permissible
Carry out according to known method of the prior art.It practice, this nanometer gold can be the gold nano of any size, shape
Grain, above-mentioned a diameter of 52.6～58.6nm gold nano grain be only used as being preferable to carry out exemplifying.
In another preference, the thickness of silicon layer is 2.5～10nm.The thickness effect laser of silicon layer is delivered to gold from shell
The efficiency of core, the most ultra-thin silicon layer is that sandwich structure Raman signal strengthens probe for fingerprint visualization successfully key.On
Stating thickness is that 2.5～10nm silicon layers are only used as being preferable to carry out exemplifying.
In another preference, the lysozyme aptamers that single sandwich structure Raman signal enhancing detecting probe surface is modified
Number is 317～351.The number of the lysozyme aptamers that detecting probe surface is modified directly affects fingerprint detection efficiency.Due to letter
Number molecule is sandwich in silicon layer, and entirely free on his silicon surface can modify the maximum amount of lysozyme aptamers, significantly improves
Recognition efficiency.The number of single detecting probe surface lysozyme aptamers is can be evaluated whether according to Lambert-Beer's law.Above-mentioned single probe
The number of surface lysozyme aptamers is 317～351 to be only used as being preferable to carry out exemplifying.
Further aspect of the present invention provides a kind of sandwich structure Raman signal for fingerprint imaging and strengthens the preparation of probe
Method, the method comprises the following steps:
(1) 613 μ L tetra-are hydrated aqueous solution of chloraurate (1% (wt/wt)) and join the round bottom filling 50mL deionized water
Flask is heated to reflux, adds 0.5mL trisodium citrate aqueous solution (1% (wt/wt)) reaction 20min after solution boiling, stop
Only heat and be cooled to room temperature, obtaining nano gold sol；
(2) ethanol solution (1mM) of 300 μ L p-Nitrobenzenethiol (pNTP) is joined nanometer gold prepared by step (1)
In colloidal sol, and stir reaction 4h, form Au@pNTP nanoparticles solution；
(3) 600 μ L 3-TSL 8330 (1mM) are joined Au@pNTP nanometer prepared by step (2)
In particle solution, it is stirred at room temperature reaction 30min, adds the sodium silicate aqueous solution (0.54% (wt/wt)) of 4.8mL,
In 90 DEG C of oil baths after stirring reaction 30min, rapidly round-bottomed flask is moved on in ice-water bath to terminate the carrying out of reaction, be cooled to room
Wen Hou, adds 300 μ L 3-TSL 8330 (1mM) and stirs reaction 15min, then dividing with ethanol and acetonitrile
Other centrifuge washing three times, finally by Au pNTP SiO2Nano-particle is dissolved in acetonitrile, standby；
(4) by the Au@pNTP@SiO prepared by step (3)2Nano-particle is molten with the acetonitrile of 3mL cyanuryl chloride (1mM)
Liquid at room temperature reacts 4h, then washs three times, washing with alcohol twice, deionized water wash twice and borate buffer with acetonitrile successively
After solution (pH=8.4) washes twice, by the Au@pNTP@SiO of activation2The borate buffer that nano-particle is scattered in 2mL again is molten
In liquid (pH=8.4), standby；
(5) by the lysozyme aptamers of 5.6nmol, (sequence is 5 '-NH2-TTT TTT ATC AGG GCT AAA GAG
TGC AGA GTT ACT TAG-3') it is dissolved in 0.4mL borate buffer solution (pH=8.4), it is then added in step (4) live
The Au@pNTP@SiO changed2In nanoparticles solution, mixture at room temperature jiggles reaction overnight, centrifugal collection bacteriolyze
The Au@pNTP@SiO that enzyme aptamers is modified2Nano-particle.
Raman signal for fingerprint imaging provided by the present invention strengthens probe and has sandwich structure: the Raman of embedding is believed
Number molecule efficiently avoid the interference of external environment condition, it is ensured that the stability of Raman signal and repeatability；Entirely free on his silicon
Layer surface can lysozyme aptamers in the maximum amount of modification, thus improve fingerprint recognition efficiency；Specific aptamers and bacteriolyze
The associativity of enzyme is good, and conjugation condition is gentle, and detection substrate need not the pre-treatment of complexity, significantly improves fingerprint imaging side
The universality of method.
Accompanying drawing explanation
Fig. 1 is the transmission of the sandwich structure Raman signal enhancing probe for fingerprint imaging of the embodiment of the present invention 1 preparation
Electromicroscopic photograph.
Fig. 2 is that the sandwich structure Raman signal of the present invention strengthens the probe high definition SERS one-tenth for grease type fingerprint ridge line
Picture.
Fig. 3 is that the sandwich structure Raman signal of the present invention strengthens the probe high definition SERS one-tenth for perspiration type fingerprint ridge line
Picture.
Detailed description of the invention
The present invention is expanded on further below in conjunction with specific embodiment.It should be understood that these embodiments be merely to illustrate the present invention and
It is not intended to limit the scope of the present invention.For indicating the experimental technique of actual conditions generally according to conventional strip in example below
Part, or according to the condition proposed by manufacturer.
In example below, lysozyme aptamers sequence used is 5 '-NH2-TTT TTT ATC AGG GCT AAA GAG
TGC AGA GTT ACT TAG-3', by Sangon Biotech (Shanghai) Co., Ltd. on behalf of synthesis.
The compound method of borate buffer solution (pH=8.4) is that 18mL borax solution (0.05mol/L) joins 22mL
Boric acid aqueous solution (0.2mol/L) shakes up, standby.
Cyanuryl chloride (2,4,6-trichloro-1,3,5-triazine) is bought in the Shanghai limited public affairs of Aladdin reagent
Department.
Embodiment 1
Sandwich structure Raman signal for fingerprint imaging strengthens the preparation of probe
(1) 613 μ L tetra-are hydrated aqueous solution of chloraurate (1% (wt/wt)) and join the round bottom filling 50mL deionized water
In flask, it is heated to reflux, adds 0.5mL trisodium citrate aqueous solution (1% (wt/wt)) reaction 20min after solution boiling, stop
Heat and be cooled to room temperature, obtaining nano gold sol.
(2) ethanol solution (1mM) of 300 μ L p-Nitrobenzenethiol (pNTP) is joined nanometer gold prepared by step (1)
In colloidal sol, and stir reaction 4h, form Au@pNTP nanoparticles solution.
(3) 600 μ L 3-TSL 8330 (1mM) are joined Au@pNTP nanometer prepared by step (2)
In particle solution, it is stirred at room temperature reaction 30min.Add the sodium silicate aqueous solution (0.54% (wt/wt)) of 4.8mL,
In 90 DEG C of oil baths after stirring reaction 30min, rapidly round-bottomed flask is moved on in ice-water bath to terminate the carrying out of reaction.It is cooled to room
Wen Hou, adds 300 μ L 3-TSL 8330 (1mM) and stirs reaction 15min, by the hydrolysis of silicone hydroxyl
Outside being exposed to by amino, thus prepare amido modified Au@pNTP@SiO2Nano-particle.It is centrifuged respectively with ethanol and acetonitrile again
Wash three times, Au pNTP SiO the most at last2Nano-particle is dissolved in acetonitrile, standby.
(4) by the Au@pNTP@SiO prepared by step (3)2Nano-particle is molten with the acetonitrile of 3mL cyanuryl chloride (1mM)
Liquid at room temperature reacts 4h.Three times, washing with alcohol twice, deionized water wash twice and borate buffer solution is washed again with acetonitrile
(pH=8.4) wash twice.Then by the Au@pNTP@SiO of activation2Nano-particle is scattered in the borate buffer solution of 2mL again
(pH=8.4) in, standby.
(5) by the lysozyme aptamers of 5.6nmol, (sequence is 5 '-NH2-TTT TTT ATC AGG GCT AAA GAG
TGC AGA GTT ACT TAG-3') it is dissolved in 0.4mL borate buffer solution (pH=8.4), it is then added in step (4) live
The Au@pNTP@SiO changed2In nanoparticles solution.Mixture at room temperature jiggles reaction overnight.Finally, lysozyme is fitted
Ligand modified Au@pNTP@SiO2Nano-particle is centrifugal to be collected, and washes twice with borate buffer solution (pH=8.4), more molten
In 2mL borate buffer solution (pH=8.4) standby.
Sandwich structure Raman signal for fingerprint imaging prepared by the present embodiment strengthens the transmission electron microscope of probe and sees figure
1, it can be seen that gold nano grain has good dispersibility, pattern is relatively uniform, and can be clear in high magnification map
Ground sees that a bright annulus is wrapped in dark golden core, illustrates that gold nano grain successfully and is completely sealed in silicon layer
In.
Embodiment 2
Sandwich structure Raman signal strengthens the probe high definition SERS imaging for grease type fingerprint
(1) collection of grease type fingerprint
Before fingerprint-collection, hands is cleaned up and after drying by volunteer with soap, with finger in forehead or wing of nose wiping again
Lightly fingerprint is pressed against the surface of the substrate of glass cleaned up.Clean substrate of glass is successively through soap by substrate
Water, ethanol and deionization clean.
(2) the high definition SERS imaging of grease type fingerprint:
After the aging 16h of fingerprint collected, sandwich structure Raman signal prepared by 200 μ L embodiments 1 strengthen probe with
It hatches 30min in the environment of room temperature (25 DEG C-27 DEG C) and humidity (relative humidity is 70%-80%).Then deionization is used
Water rinses the probe removing non-specific adsorption, and rear use Horiba XploRA Confocal laser-scanning microscopy instrument to be dried draws
Graceful light spectrum image-forming, a length of 635nm of Raman spectrum excitation light wave, acquisition time is 0.1s.
SERS imaging results sees Fig. 2.
Embodiment 3:
Sandwich structure Raman signal strengthens the probe high definition SERS imaging for perspiration type fingerprint
(1) collection of perspiration type fingerprint
Before fingerprint-collection, hands is cleaned up with soap and after drying, puts on PE glove and seal antiperspirant 10min, then by volunteer
Pressing lightly on 20s at clean glass basic surface, clean substrate of glass is successively through suds, ethanol with go by substrate
Ion Cleaning.
(2) the high definition SERS imaging of perspiration type fingerprint, step is with the step (2) of above-described embodiment 2.
SERS imaging results sees Fig. 3.
Embodiments above illustrates that the sandwich structure Raman signal for fingerprint imaging of the present invention strengthens probe
Preparation method and using effect thereof, but they are only presently preferred embodiments of the present invention, are not limited to the scope of the present invention.
The above embodiment of the present invention can also make a variety of changes, the most every claims according to the present patent application and description
Change simple, equivalent that content is made and modification, fall within the claims of patent of the present invention.The present invention is the most detailed
Describe is routine techniques content.
Claims (10)
1. the sandwich structure Raman signal for fingerprint imaging strengthens probe, it is characterised in that described probe is with gold nano
Granule is core, and silicon dioxide is shell, and signaling molecule is sandwich between described core and described shell, described shell surface be modified with for
The lysozyme aptamers of fingerprint recognition.
Sandwich structure Raman signal for fingerprint imaging the most according to claim 1 strengthens probe, it is characterised in that institute
Stating signaling molecule is p-Nitrobenzenethiol, mercaptopyridine or mercaptobenzoic acid.
Sandwich structure Raman signal for fingerprint imaging the most according to claim 1 strengthens probe, it is characterised in that institute
The sequence stating lysozyme aptamers is 5 '-NH2-TTT TTT ATC AGG GCT AAA GAG TGC AGA GTT ACT TAG-
3'。
Sandwich structure Raman signal for fingerprint imaging the most according to claim 1 strengthens probe, it is characterised in that institute
State a diameter of the 52.6～58.6nm of gold nano grain.
Sandwich structure Raman signal for fingerprint imaging the most according to claim 1 strengthens probe, it is characterised in that institute
The thickness stating silicon layer is 2.5～10nm.
6. the sandwich structure Raman signal for fingerprint imaging strengthens a preparation method for probe, and the method includes following step
Rapid: (1) prepares gold nano grain；(2) by S-Au key, signaling molecule is connected on described gold nano grain；(3) silicon is passed through
Acid sodium hydrolysis superscribes silicon layer；(4) by amido modified lysozyme aptamers function being existed at silicon surface formation triazine radical
Its surface.
Sandwich structure Raman signal the most according to claim 6 strengthens the preparation method of probe, it is characterised in that described letter
Number molecule is p-Nitrobenzenethiol, mercaptopyridine or mercaptobenzoic acid.
Sandwich structure Raman signal for fingerprint imaging the most according to claim 6 strengthens probe, it is characterised in that institute
The sequence stating lysozyme aptamers is 5 '-NH2-TTT TTT ATC AGG GCT AAA GAG TGC AGA GTT ACT TAG-
3'。
Sandwich structure Raman signal for fingerprint imaging the most according to claim 6 strengthens probe, it is characterised in that institute
State a diameter of the 52.6～58.6nm of gold nano grain.
10. the sandwich structure Raman signal for fingerprint imaging strengthens a preparation method for probe, and the method includes following step
Rapid:
(1) 613 μ L tetra-are hydrated aqueous solution of chloraurate (1% (wt/wt)) and join the round-bottomed flask filling 50mL deionized water
In be heated to reflux, solution boiling after add 0.5mL trisodium citrate aqueous solution (1% (wt/wt)) reaction 20min, stop adding
Heat is also cooled to room temperature, obtains nano gold sol；
(2) ethanol solution (1mM) of 300 μ L p-Nitrobenzenethiol (pNTP) is joined nano gold sol prepared by step (1)
In, and stir reaction 4h, form Au@pNTP nanoparticles solution；
(3) 600 μ L 3-TSL 8330 (1mM) are joined Au@pNTP nano-particle prepared by step (2)
In solution, after being stirred at room temperature reaction 30min, add the sodium silicate aqueous solution (0.54% (wt/wt)) of 4.8mL, at 90 DEG C
In oil bath after stirring reaction 30min, rapidly round-bottomed flask is moved on in ice-water bath to terminate the carrying out of reaction, be cooled to room temperature
After, add 300 μ L 3-TSL 8330 (1mM) and stir reaction 15min, then distinguishing with ethanol and acetonitrile
Centrifuge washing three times, finally by Au@pNTP@SiO2Nano-particle is dissolved in acetonitrile, standby；
(4) by the Au@pNTP@SiO prepared by step (3)2The acetonitrile solution of nano-particle and 3mL cyanuryl chloride (1mM) is in room
React 4h under temperature, then wash three times, washing with alcohol twice, deionized water wash twice and borate buffer solution with acetonitrile successively
(pH=8.4) wash twice, then by the Au@pNTP@SiO of activation2Nano-particle is scattered in the borate buffer solution of 2mL again
(pH=8.4) in, standby；
(5) by the lysozyme aptamers of 5.6nmol, (its sequence is 5 '-NH2-TTT TTT ATC AGG GCT AAA GAG TGC
AGA GTT ACT TAG-3') it is dissolved in 0.4mL borate buffer solution (pH=8.4), it is then added to activation in step (4)
Au@pNTP@SiO2In nanoparticles solution, mixture at room temperature jiggles reaction overnight, and centrifugal collection lysozyme is adaptive
The Au@pNTP@SiO that body is modified2Nano-particle.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110031446A (en) * | 2019-04-28 | 2019-07-19 | 河南大学 | The method for detecting photic nanoparticles in polymers spatial distribution using Raman image technology |
| CN113125410A (en) * | 2021-04-19 | 2021-07-16 | 湖南大学 | Multi-material-substrate universal latent fingerprint imaging and residue detection system based on gold-graphite nanocapsule particles and imaging method thereof |
| CN114309591A (en) * | 2021-12-30 | 2022-04-12 | 杭州电子科技大学 | Self-assembly method for coating thin silicon dioxide gold nanorods |
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