CN105823770B - A kind of Raman labels probe of no optical interference and its preparation method and application - Google Patents
A kind of Raman labels probe of no optical interference and its preparation method and application Download PDFInfo
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Abstract
The present invention with to a series of Raman scattering peak positions of structure design based on sulfydryl phenylacetylene in 1,800 2400 cm‑1Wave number area signaling molecule, select enhancing substrate of the gold, silver nano-particle as Raman signal, alkynes signaling molecule with sulfydryl is self-assembled to enhancing substrate surface and SERS signal is optimized using thioacetic acid, then it selects light transmission, hydrophilic and lapping rich in active reactive group to carry out probe encapsulation, finally grafts specific biological targeting functional molecular in lapping outer layer.The optic response of this probe Raman scattering is strong, the unimodal transmitting in narrowband, without optics background interference, it is significant in the bio-imaging field that multicomponent marks simultaneously.The technology may extend to using bandpass filter as light-splitting device, with photomultiplier（PMT）Or avalanche photodide（APD）For the design and development of the ultrafast Raman image instrument of no-raster of detection device, overcomes Raman image point by point scanning, is imaged slow technical bottleneck, the market vacancy of optical imagery quasi-instrument will be filled up.
Description
Technical field
The invention belongs to bio-imaging technical fields, are related to a kind of surface enhanced drawing of novel alkynes coding, biological targeting
The preparation method of graceful scattering (Surface-enhanced Raman scattering, SERS) active nano label probe.It is heavier
It wants, is related to using bandpass filter as light-splitting device, is inspection with photomultiplier (PMT) or avalanche photodide (APD)
Survey the design and development of the ultrafast Raman image instrument of no-raster of device.
Background technology
Bio-imaging (such as living cells imaging) technology allows scientist in real time and dynamic observation organism internal structure
And physiology course, thoroughly reform the mode of biologist postgraduate's object movement mechanism.In numerous bio-imaging technologies,
Fluorescent marker imaging applications are the most extensively and ripe.It is using the light for caning absorb specific wavelength and launches the longer light of wavelength
The signal probe of (fluorescence), for example, small molecule organic dyestuff or fluorescin it is specific in biosystem specifically to mark
Component, and the distribution situation of target organism substance is obtained by the positioning of its fluorescence signal, greatly improve people's exploration
The ability of organism internal structure and process.However, this imaging technique is obviously limited by the inherent shortcoming of fluorescent spectrometry, i.e.,
Traditional fluorescent dye transmitting peak width (about 50nm) is wide to be caused signal overlap when polychrome (or multicomponent) label and be difficult to area
Divide, discrimination.The progress of technology of quantum dots in recent years overcomes the inherent shortcoming of above-mentioned traditional organic fluorescent dye and fluorescin,
Its emission peak is narrow and symmetrical, overlapped small and its emission peak wavelength can be made from it material and grain size fine-tunes etc.
Good characteristic by cell marking and imaging research favor, but its potential bio-toxicity make its be applied to living imaging one
A little further investigation difficulties have new progress.
Surface enhanced Raman scattering (Surface-enhanced Raman scattering, SERS) technology is removed with non-
Destructive, EO-1 hyperion specificity and not by outside the advantages of the normal Raman spectroscopies such as water interference, also have lower detection limit with more
High sensitivity (ability for having Single Molecule Detection) especially intervenes the situation of biomedical sector in nanotechnology advantage
Under, various gold (silver) nano particles with surface enhanced Raman scattering effect be widely used in biological sample analysis detection with
Bio-imaging is studied.In order to improve the sensitivity of Raman detection, people have carried out greatly the selection and preparation of SERS base materials
Quantifier elimination.However, the key factor as another decision probe quality, the selection and design of signaling molecule do not cause but
The enough attention of people.The it is proposed of novel signaling molecule is usually required by reasonably designing, careful screening and system
Characterization, compared to then undoubtedly seeming excessively cumbersome for the SERS probes of selection " ready-made ", to cause nowadays about letter
The work of number MOLECULE DESIGN is fewer and fewer.But in complicated research system, the random selection of SERS probe molecules but may be used
It can become the obstacle that research further develops, because disturbing molecule or detection molecules are easily sent out with traditional SERS signal molecule
The phenomenon that raw spectra overlapping, this will seriously affect the accuracy of result.With multiple target simultaneously detection concept proposition, this
Puzzlement even can extend between all kinds of SERS probe molecules, at this moment classical signal molecule selection difficulty will further by
Amplification.In face of such predicament, there is an urgent need to a large amount of mutually non-overlapping novel signaling molecules for the anti-dry of complex system by people
It disturbs, multiobjective analysis detection.
The raman characteristic peak of alkynyl is in 2120cm-1Near, it is not have in the intracellular common interfering substance of this SPECTRAL REGION
There is Raman response, so the biological metabolism molecule that people mark alkynes in many researchs in recent years, such as DNA, RNA, egg
White matter, lipid etc. are imaged for biomarker and achieve good effect；In addition, the Raman shift of alkynyl and intensity can because
The difference for the substituent group being attached thereto and occur obviously to deviate, or even be only by the carbon atom of alkynyl12C is changed to its same position
Element13C, Raman shift just will produce the offset of 100 wave numbers, this 2 points of derivatives predictive of alkynes are extremely suitable jamproof
Multiobjective analysis detects.But the extensive use of alkynes class signaling molecule is but still very weak by spontaneous Raman scattering signal strength
The problem of seriously limit, in order to detect extremely faint alkynyl Raman scattering signal, people have to improve the time for exposure or
Using high intensity laser beam, thus but the damage of sample may be caused；It also has tried to use stimulated Raman scattering in recent years
(Stimulated Raman scattering, SRS) technology carries out correlative study, but expensive instrument price allow people hope and
While raw fear, in the awkward general instrument for also making it that can not become bio-imaging field that multiple target is imaged simultaneously
Device.In conjunction with the more SERS technologies of recent researches, it provides 106~1012Signal enhancing again will be theoretically an improvement
The weaker desirable route of alkynyl spontaneous Raman scattering signal.
Invention content
The purpose of the present invention is to provide the Surface enhanced Raman scattering of a kind of novel alkynes coding, biological targeting
The preparation method of (Surface-enhanced Raman scattering, SERS) active nano label probe, it is prior
It is to develop a kind of high brightness, polychrome output, optical stabilization and glitch-free novel SERS imaging techniques.The technology (including probe
Product) it can realize extensive bio-imaging application on conventional commercialization confocal spectrometer；Meanwhile it can also be used for band
Pass filter is light-splitting device, is surpassed with the no-raster that photomultiplier (PMT) or avalanche photodide (APD) are detection device
The design and development of fast Raman image instrument.
Technical solution provided by the invention is as follows：
A kind of preparation method of the Raman labels probe of no optical interference, includes the following steps：Raman scattering peak position is existed
1800-2400cm-1Wave number area, the unimodal signaling molecule in narrowband that bandwidth is 1-2nm self-assemble to increasing by Au-S keys or Ag-S keys
Then the surface at strong basis bottom uses the hydrophilic case material of light transmission to carry out probe encapsulation to get to the Raman mark of no optical interference
Remember probe；
The unimodal Raman signal molecule in the narrowband has structure shown in general formula I：
Wherein, R1、R2、R3、R4For-NO2、-CH3Or H, R5For H or-Si (CH3)3、-C2H5、-CH2CH2CH2OH, phenyl or
Alkynyl；
The enhancing substrate is the nano-particle of grain size 30-60nm, and the nano-particle is (i) nanogold, (ii)
Nano silver or (iii) are using nanogold or nano silver as the core-shell structure nanometer particle of shell；
The case material is one kind in biomolecule, polymer, silica.
Further, R5For H the unimodal signaling molecule in narrowband self-assemble to enhancing substrate surface after, with mercaptan carboxylic acid couple
The unimodal signaling molecule in narrowband and the connection type of enhancing substrate optimize, and the case material for then using light transmission hydrophilic is visited
Needle encapsulates.
Still further, biological targeting molecule is grafted after probe is encapsulated on case material.
The unimodal Raman signal molecule in the narrowband is
The mercaptan carboxylic acid is one or more of thioacetic acid, mercaptopropionic acid, mercaptobutyric acid；The biological target
It is one kind in antibody, ligand, polypeptide to molecule.
The antibody is luteinising hormone-releasing hormo, and the ligand is folic acid, and the polypeptide is cell-penetrating
Peptide or nuclear location peptide.
The biological targeting molecule is grafted onto detecting probe surface by being reacted with EDC-NHS or glutaraldehyde cross-linking.
A kind of Raman labels probe of no optical interference, by the preparation side of the Raman labels probe of the no optical interference
Method is prepared.
The application of the Raman labels probe in bio-imaging without optical interference.
A kind of highly sensitive ultrafast Raman image instrument is detection object with the Raman labels probe of the no optical interference,
On the basis of conventional light microscope, using laser as excitation light source, high-precision quick scanning sample stage is configured, with ultra-narrow
The unimodal superpower Raman scattering signal in narrowband that probe is extracted with bandpass filter is detection with overdelicate avalanche photodide
Element by the photon signal of the point-by-point acquisition probe of the two-dimensional scan of sample stage and makees Pseudo Col ored Image.
This quick, sensitive and high-space resolution Raman image instrument is with the Wavelength calibration aberration of bandpass filter, with photon
The intensity calibration coloration of signal.
Multiple targets are marked in the Raman labels probe of the no optical interference, use the two-dimentional automatically scanning of band
The Raman spectrometer of platform rapidly acquires spectrum, and then fakes to the unimodal superpower Raman scattering signal in probe narrowband point by point
Color Image Processing, the displacement of scattering peak is for demarcating aberration, and the intensity of scattering peak is for demarcating coloration；The Quick Acquisition
The condition of spectrum is：Time for exposure is 1s, and wave-length coverage is Δ 50cm-1；The target is cell, tissue or organism.
The principle of the present invention is：
1. with to structure based on sulfydryl phenylacetylene, change alkynyl Raman scattering by replacing alkynyl terminal substituent group
The displacement of signal can carry out one according to Density function theory to the relationship of structure-Raman shift of alkynes class signaling molecule
Serial anticipation and theoretical research filter out the unimodal molecular structure of different narrow band.
2. synthesizing the gold, silver and its core-shell structure nanometer particle of different shape, size, the high active nanometer of enhancing is filtered out
Particle enhances substrate as SERS, by alkynes signaling molecule by Au-S or Ag-S keys self-assembled monolayer in enhancing substrate surface.
3. the connection side to signaling molecule and enhancing substrate such as thioacetic acid can be used in the self assembling process of signaling molecule
Formula optimizes, and to eliminate due to Interference Peaks caused by spatial orientation difference, ensures in 1800~2400cm-1In spectrum range
The unimodal output mode in narrowband.
4. using large biological molecule, polymer or titanium dioxide silicon material as protection shell, encapsulation self assembly has signaling molecule expensive
The probes parent such as metal nanoparticle, outer surface is provided is conducive to transferring for subsequent bio functional molecular rich in reaction active groups
It connects.
5. with ligand (such as folic acid), antibody (such as luteinising hormone-releasing hormo), polypeptide, (such as cell-penetrating peptides are appraised and decided
Position peptide etc.) etc. biologically functional molecules pass through the cross-linking reactions such as EDC-NHS or glutaraldehyde to detecting probe surface, you can have it is special raw
The SERS image probes of object target function.
Relative to existing SERS image probes, the present invention has the following advantages and beneficial effect：
1. the probe is in 1800-2400cm-1Spectral regions emit narrowband (1-2nm) unimodal superpower Raman scattering signal, energy
It is enough effectively avoid cellular endogenous substance lower wave number region (<1800cm-1) intrinsic Raman signal and fluorescence signal interference；It should
The alkynyl Raman shift of the used signaling molecule of probe can be by changing to alkynyl end in sulfydryl phenylacetylene agent structure
Substituent group is adjusted, and can effectively avoid the problem that overlapping each other between marking signal in polychrome imaging applications；The probe is believed
Number intensity is big, optical stabilization, can effectively shorten the bio-imaging time using normal Raman spectrometer, the unimodal output mould in narrowband
Formula can be used for the design and development of the ultrafast Raman image instrument of no-raster.
2. the optic response of the probe Raman scattering is strong, the unimodal transmitting in narrowband, without optics background interference, multicomponent simultaneously
The bio-imaging field of label is significant.The technology may extend to using bandpass filter as light-splitting device, with photomultiplier
(PMT) or avalanche photodide (APD) be detect device the ultrafast Raman image instrument of no-raster design and development, overcome
The slow technical bottleneck of Raman image point by point scanning, imaging, will fill up the market vacancy of optical imagery quasi-instrument.
Description of the drawings
Fig. 1 is the principle schematic that thioacetic acid improves the connection type of signaling molecule and enhancing substrate.
Fig. 2 is the Raman shift spectrum comparison diagram of tri- kinds of signaling molecules of B1, B2, B3.
Fig. 3 is the spectral intensity comparison diagram that thioacetic acid improves the connection type of signaling molecule and enhancing substrate.
Fig. 4 is cell imaging of the SERS probes of B3 signaling molecules preparation from pollen cell endogenous substance at different wave numbers
Comparison diagram；Wherein, Fig. 4 (A) is 1580cm-1Cell imaging figure, Fig. 4 (B) be 2212cm-1Cell imaging figure.
Fig. 5 is the attached cell image of SERS probes prepared by tri- kinds of signaling molecules of B1, B2, B3.
Fig. 6 is spectrogram of the tri- kinds of SERS probes of B1, B2, B3 in same cell.
Fig. 7 is cell imaging figure of the tri- kinds of SERS probes of B1, B2, B3 in same cell.
Specific implementation mode
With reference to example, the present invention is described in further detail, and the advantage of this method is proved with example, but this
The embodiment of method is without being limited thereto.
Embodiment 1：The preparation of the unimodal Raman signal molecule in narrowband
The unimodal Raman signal MOLECULE DESIGN in alkynes class narrowband：The present invention is with to structure based on sulfydryl phenylacetylene, by replacing
Alkynyl terminal substituent group is changed to change the displacement of alkynyl Raman scattering signal, and using Density function theory to signal point
The relationship of structure-Raman shift of son carries out a series of predictions and theoretical research, has obtained the chemical constitution in following table and its drawing
Graceful displacement, it was demonstrated that can effectively adjust the unimodal signaling molecule in narrowband when changing group in the alkynyl end to sulfydryl phenylacetylene
The displacement of Raman signal.
Signaling molecule spectrum optimizes：It is enhancing substrate with the silver-colored gold-covered nano particle of certain size, using 532nm to mercapto
The Raman spectrum of base phenylacetylene molecule optimizes, and finds when gold nanoparticle is added to sulfydryl phenylacetylene, can be in 1972cm-1
And 2105cm-1Place generates a Raman signal respectively, is confirmed this is because signaling molecule and increasing by Density function theory
It, can be to signaling molecule and enhancing substrate when finding to be added thioacetic acid by research caused by the spatial orientation difference at strong basis bottom
Connection type improved, result schematic diagram is as shown in Figure 1.
Signaling molecule SERS spectra：According to signaling molecule spectrum optimum results, select the unimodal signaling molecule B1 in three kinds of narrowbands,
For B2, B3, obtained practical SERS spectra is as shown in Fig. 2, the alkynyl Raman shift of three is located at 2105cm-1、
2158cm-1、2212cm-1Place, can be effectively prevented from mutual interference.
Embodiment 2
(1) AuNPs- mark molecules：Three parts of 10mL silver gold-covered nanos colloidal sol is taken, it is a concentration of that 10 μ L are added thereto respectively
The tetrahydrofuran solution of narrowband unimodal signaling molecule B1, B2, B3 of 1mM.
(2) thioacetic acid optimizes：The water of the thioacetic acid of 100 a concentration of 10mmol/L of μ L is added after above-mentioned solution left standstill 3h
Solution.(this step is only needed when the alkynyl end of signaling molecule does not replace, for example using B1 as signaling molecule).
(3) shell is protected：Polycyclic aromatic hydrocarbon (PAH) aqueous solution that a concentration of 0.1wt of 1mL ‰ are continuously added after 3h is stood again.
(4) biologically functional molecule connects：The 1- (3- dimethylamino-propyls)-of 10 a concentration of 10mM of μ L is added after 12h simultaneously
N-hydroxysuccinimide (NHS) water of 3- ethyl-carbodiimide hydrochlorides (EDCHCl) aqueous solution and 20 a concentration of 1mM of μ L
Solution stands activated carboxyl, folic acid (FA), the metakentrin of 20 a concentration of 1mM of μ L is added after 1h into three parts of solution respectively
Releasing hormone (LHRH), cell-penetrating peptides (CALNNR8) solution.
(5) it is about 10 to be collected by centrifugation and be resuspended to particle concentration after standing a night12A/mL, is put into refrigerator and saves backup.
There is the SERS probes of wherein FA and LHRH modifications cell membrane to target ability, CALNNR8The SERS probes of modification have endoplasmic reticulum
Targeting ability.
Using B1 as signaling molecule, optimized using different amounts of thioacetic acid：Take the B1-AuNPs nanometers of three parts of 10mL
Particle is separately added into the aqueous solution of the thioacetic acid of 0,1,10,100 a concentration of 10mmol/L of μ L thereto, after standing three hours
The SERS spectra measured, as shown in Figure 3.
Embodiment 3
It is about 10 to take 10mL particle concentrations12The silver-colored gold-covered nano colloidal sol of a/mL is added 20 a concentration of 1mM's of μ L thereto
B3 signaling molecules stand polycyclic aromatic hydrocarbon (PAH) aqueous solution for continuously adding a concentration of 0.1wt of 1mL ‰ after 3h, continue to stand 12h
SERS probes are made afterwards.
It takes 10mL is evenly dispersed to have lily powder aqueous solution, and 1mLSERS probes, the mixing on shaking table is added thereto
Pollen solution is lyophilized to remove moisture afterwards.
The sucrose solution of compounding high concentration is simultaneously added dropwise on a small quantity on glass slide, dries the sucrose that rear surface layer forms homogeneous transparent
Layer.It takes a small amount of pollen particles to trickle down on sucrose layer and gently blows to apply pressure, pollen is made to be firmly adhered to sucrose table
Raman image experiment can be carried out behind face.
With the flavonoid class substance in pollen in 1580cm-1Raman signal be Interference Peaks carry out confirmatory experiment.It can send out
Existing, alkynyl signaling molecule is in 1580cm-1The Raman signal in lower wave number region is easy to that spectrum weight occurs with the Interference Peaks in pollen
Folded phenomenon (see Fig. 4 (A)), this will produce very big interference to final result, and use 2212cm-1Marker peak then can effectively avoid
Problems (see Fig. 4 (B)).Confirm alkynyl SERS probes in cell imaging research to the anti-interference energy of cellular endogenous substance
Power.
Embodiment 4
It is about 10 to take 10mL particle concentrations1220 μ L concentration are added in three parts of the silver-colored gold-covered nano colloidal sol of a/mL thereto respectively
For the tetrahydrofuran solution of narrowband unimodal signaling molecule B1, B2, B3 of 1mM.It is a concentration of that 1mL is continuously added after standing 3h
Mercapto-polyglycol (HS-PEG) aqueous solution of 0.1wt ‰ can be prepared by corresponding SERS probes after standing 6h.
It is put into the glass cover-slip (passing through high-temperature sterilization) of a diameter of 20mm in batch cultur ware (35mm), takes thin
Finely dispersed cell suspending liquid (300~500 μ L) drops evenly to coverslip surface after the final step dilution of born of the same parents' passage.
After standing 2-4h in incubator, cell is substantially adherent, adds 2.5mL fresh cell mediums, waits for 4h.
The attached cell for taking three parts of above methods to prepare, and it is separately added into three kinds of SERS probes thereto, in the incubator
Continue to take out after co-culturing 12h, coverslip gently rinsed using PBS buffer solutions, removing be adsorbed in cell membrane surface and not into
Enter the SERS probes in cell, imaging experiment can be carried out later.
As shown in figure 5, three kinds of SERS probes present good distribution situation in the cell, and each channel it
Between entirely without influence, it was demonstrated that three kinds of alkynes SERS probes will not interfere between each other.
Embodiment 5
It is about 10 to take 10mL particle concentrations1220 μ L concentration are added in three parts of the silver-colored gold-covered nano colloidal sol of a/mL thereto respectively
For the tetrahydrofuran solution of narrowband unimodal signaling molecule B1, B2, B3 of 1mM.It is a concentration of that 1mL is continuously added after standing 3h
The polypropylene amine aqueous solution of 0.1wt ‰.1- (3- dimethylamino-propyls) -3- second of 10 a concentration of 10mM of μ L is added after 12h simultaneously
N-hydroxysuccinimide (NHS) aqueous solution of base carbodiimide hydrochloride (EDCHCl) aqueous solution and 20 a concentration of 1mM of μ L.
The folic acid (FA) of 20 a concentration of 1mM of μ L is added after standing activated carboxyl 1h into three parts of probes respectively, metakentrin release swashs
Plain (LHRH), cell-penetrating peptides (CALNNR8) solution.It is collected by centrifugation after standing a night and can be prepared by three kinds of corresponding SERS spies
Needle.
It is put into the glass cover-slip (passing through high-temperature sterilization) of a diameter of 20mm in batch cultur ware (35mm), takes thin
Finely dispersed cell suspending liquid (300~500 μ L) drops evenly to coverslip surface after born of the same parents pass on final step dilution.It is training
It supports after standing 2-4h in case, cell is substantially adherent, adds 2.5mL fresh cell mediums, waits for 4h.
The completely adherent rear addition of cell has the SERS probes (B3-CLANNR of endoplasmic reticulum targeting ability8- NPs) 200 μ L are simultaneously
Gently mixing.Continue to take out after co-culturing 12h in the incubator, gently rinses coverslip using PBS buffer solutions, remove absorption
The endoplasmic reticulum targeted probes in cell are not entered in cell membrane surface, are then added in 3mL fresh cultures to small culture dish,
And two kinds of cell membranes are added and target SERS probes (B1-FA-NPs and OPE1-LHRH-NPs) each 200 μ L.Continue after cultivating 2-4h
It can carry out SERS imaging research.
Fig. 6 is the more typical SERS spectra measured in cell, it can be found that the signal of three kinds of mark molecules can
It most clearly distinguishes, and will not be interfered completely by cellular endogenous substance.Fig. 7 is that three kinds of probes pass through in same cell
The imaging results that detection actually obtains are crossed, three kinds of SERS probes realize according to the effect of biologically functional molecule selectively targeted
As a result.
Claims (9)
1. a kind of preparation method of the Raman labels probe of no optical interference, which is characterized in that include the following steps：Raman is dissipated
Peak position is penetrated in 1800-2400cm-1Wave number area, the unimodal signaling molecule in narrowband that bandwidth is 1-2nm by Au-S keys or Ag-S keys from
It is assembled into the surface of enhancing substrate, the hydrophilic case material of light transmission is then used to carry out probe encapsulation to get to no optical interference
Raman labels probe；
The unimodal Raman signal molecule in the narrowband is Or the compound with structure shown in general formula I；
Wherein, R1、R2、R3、R4For-NO2、-CH3Or H, R5For H or-Si (CH3)3、-C2H5、-CH2CH2CH2OH, phenyl or alkynyl；
The enhancing substrate is the nano-particle of grain size 30-60nm, and the nano-particle is (i) nanogold, (ii) nanometer
Silver or (iii) are using nanogold or nano silver as the core-shell structure nanometer particle of shell；
The case material is one kind in biomolecule, polymer, silica.
2. the preparation method of the Raman labels probe of no optical interference according to claim 1, it is characterised in that：R5For H's
After the unimodal signaling molecule in narrowband self-assembles to the surface of enhancing substrate, with mercaptan carboxylic acid to the unimodal signaling molecule in narrowband and enhancing base
The connection type at bottom optimizes, and the case material for then using light transmission hydrophilic carries out probe encapsulation.
3. the preparation method of the Raman labels probe of no optical interference according to claim 2, it is characterised in that：By probe
Biological targeting molecule is grafted after encapsulation on case material.
4. the preparation method of the Raman labels probe of no optical interference according to claim 3, it is characterised in that：Described
The unimodal Raman signal molecule in narrowband is
5. the preparation method of the Raman labels probe of no optical interference according to claim 3, it is characterised in that：Described
Mercaptan carboxylic acid is one or more of thioacetic acid, mercaptopropionic acid, mercaptobutyric acid；The biological targeting molecule be antibody,
One kind in ligand, polypeptide.
6. the preparation method of the Raman labels probe of no optical interference according to claim 5, it is characterised in that：Described
Antibody is luteinising hormone-releasing hormo, and the ligand is folic acid, and the polypeptide is cell-penetrating peptides or nuclear location peptide.
7. the preparation method of the Raman labels probe of no optical interference according to claim 5, it is characterised in that：Described
Biological targeting molecule is grafted onto detecting probe surface by being reacted with EDC-NHS or glutaraldehyde cross-linking.
8. a kind of Raman labels probe of no optical interference, it is characterised in that：By claim 1-7 any one of them without optics
The preparation method of the Raman labels probe of interference is prepared.
9. application of the Raman labels probe of no optical interference according to any one of claims 8 in bio-imaging.
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| CN108037108B (en) * | 2017-12-05 | 2019-12-10 | 武汉大学 | method for measuring content of thiocyanate ions in actual sample |
| CN108562569B (en) * | 2018-06-04 | 2021-01-01 | 中国人民解放军第二军医大学 | Circulating tumor cell detection method based on surface-enhanced Raman spectrum probe |
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| CN102323312A (en) * | 2011-09-21 | 2012-01-18 | 福州大学 | Detection method and device for portable rapid visual detection of copper ion content |
| CN103674925B (en) * | 2012-09-14 | 2016-08-10 | 北京大学 | 4-mercaptophenyl boronic acid modify golden nanometer particle and with its detection cell surface sugar labelling method |
| CN103454268B (en) * | 2013-04-19 | 2016-02-24 | 南京工业大学 | A kind of reducing sugar quantitative detecting method based on click-reaction |
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