CN110231316A - The preparation of bimodal nano-probe and its label and imaging to mescenchymal stem cell - Google Patents
The preparation of bimodal nano-probe and its label and imaging to mescenchymal stem cell Download PDFInfo
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- 230000002902 bimodal effect Effects 0.000 title claims abstract description 25
- 239000000523 sample Substances 0.000 title claims abstract description 21
- 238000003384 imaging method Methods 0.000 title claims abstract description 19
- 238000002360 preparation method Methods 0.000 title claims description 9
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 47
- 229910052681 coesite Inorganic materials 0.000 claims abstract description 18
- 229910052906 cristobalite Inorganic materials 0.000 claims abstract description 18
- 229910052682 stishovite Inorganic materials 0.000 claims abstract description 18
- 229910052905 tridymite Inorganic materials 0.000 claims abstract description 18
- 239000000377 silicon dioxide Substances 0.000 claims abstract description 14
- 238000000034 method Methods 0.000 claims abstract description 13
- 230000005311 nuclear magnetism Effects 0.000 claims abstract description 10
- 238000001354 calcination Methods 0.000 claims abstract description 5
- 239000008367 deionised water Substances 0.000 claims abstract description 5
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- 238000001035 drying Methods 0.000 claims abstract description 4
- 230000008569 process Effects 0.000 claims abstract description 3
- 239000000243 solution Substances 0.000 claims description 36
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 21
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 claims description 13
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- BOTDANWDWHJENH-UHFFFAOYSA-N Tetraethyl orthosilicate Chemical compound CCO[Si](OCC)(OCC)OCC BOTDANWDWHJENH-UHFFFAOYSA-N 0.000 claims description 3
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- GWEVSGVZZGPLCZ-UHFFFAOYSA-N Titan oxide Chemical compound O=[Ti]=O GWEVSGVZZGPLCZ-UHFFFAOYSA-N 0.000 description 2
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- WZZBNLYBHUDSHF-DHLKQENFSA-N 1-[(3s,4s)-4-[8-(2-chloro-4-pyrimidin-2-yloxyphenyl)-7-fluoro-2-methylimidazo[4,5-c]quinolin-1-yl]-3-fluoropiperidin-1-yl]-2-hydroxyethanone Chemical compound CC1=NC2=CN=C3C=C(F)C(C=4C(=CC(OC=5N=CC=CN=5)=CC=4)Cl)=CC3=C2N1[C@H]1CCN(C(=O)CO)C[C@@H]1F WZZBNLYBHUDSHF-DHLKQENFSA-N 0.000 description 1
- NPRFXRHWTRERQD-UHFFFAOYSA-N 1-methyl-1,2,3,4,5-pentakis-phenylsilole Chemical compound C=1C=CC=CC=1[Si]1(C)C(C=2C=CC=CC=2)=C(C=2C=CC=CC=2)C(C=2C=CC=CC=2)=C1C1=CC=CC=C1 NPRFXRHWTRERQD-UHFFFAOYSA-N 0.000 description 1
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- 239000002202 Polyethylene glycol Substances 0.000 description 1
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- UIWYJDYFSGRHKR-UHFFFAOYSA-N gadolinium atom Chemical compound [Gd] UIWYJDYFSGRHKR-UHFFFAOYSA-N 0.000 description 1
- XWFVFZQEDMDSET-UHFFFAOYSA-N gadolinium(3+);trinitrate;hexahydrate Chemical compound O.O.O.O.O.O.[Gd+3].[O-][N+]([O-])=O.[O-][N+]([O-])=O.[O-][N+]([O-])=O XWFVFZQEDMDSET-UHFFFAOYSA-N 0.000 description 1
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- 238000002372 labelling Methods 0.000 description 1
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- 238000002595 magnetic resonance imaging Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 210000002901 mesenchymal stem cell Anatomy 0.000 description 1
- 210000003716 mesoderm Anatomy 0.000 description 1
- XZWYZXLIPXDOLR-UHFFFAOYSA-N metformin Chemical compound CN(C)C(=N)NC(N)=N XZWYZXLIPXDOLR-UHFFFAOYSA-N 0.000 description 1
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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
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N21/6456—Spatial resolved fluorescence measurements; Imaging
- G01N21/6458—Fluorescence microscopy
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N24/00—Investigating or analyzing materials by the use of nuclear magnetic resonance, electron paramagnetic resonance or other spin effects
- G01N24/08—Investigating or analyzing materials by the use of nuclear magnetic resonance, electron paramagnetic resonance or other spin effects by using nuclear magnetic resonance
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- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- High Energy & Nuclear Physics (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
The present invention relates to field of biotechnology, mesoporous silicon oxide is sintered into fine silica end, by Gd (NO by bimodal nano-probe synthetic method3)3·6H2O dissolution forms Gd (NO in deionized water3)3Solution, then by Gd (NO3)3Solution is added dropwise in fine silica end, and dropwise addition process is stirred, then drying and calcination, obtains Gd@SiO2Compound；The method of the present invention prepares AIE-Gd@SiO2Nano-complex has good biocompatibility and hypotoxicity, can be used as nano-probe label mescenchymal stem cell, while fluorescence and the imaging of nuclear-magnetism bimodal, the experimental study applied to stem cell may be implemented.
Description
Technical field
The present invention relates to field of biotechnology, specifically prepare bimodal nano-probe AIE-Gd@SiO2, it is used for marking
Remember mescenchymal stem cell, realizes and the fluorescence and nuclear-magnetism bimodal of stem cell are imaged.
Background technique
Mescenchymal stem cell (MSCs) is a kind of from the mesoblastic multipotential stem cell of mesoderm growing early stage, has multidirectional point
Change potential, improve and rehabilitate the biological characteristics such as injury tissue, immunoregulation and hematopoiesis support function.In suitable environment it
Orientable be divided into osteoblast, cartilage cell, cardiac muscle cell and fat cell etc..Lot of experiments shows stem cell
It is divided between osteoblast and fat cell that there is significantly plasticity, and the two can mutually convert.This shows
It by inhibiting mescenchymal stem cell lipoblast to break up and increase its Osteoblast Differentiation, is prevented or treats due to bone content reduces
The important measures of caused bone disease, have attracted special attention in bone tissue engineer.21st century is in stem-cell research
Aspect has obtained numerous scientific discoveries, us is made to expand the understanding to cell biology.Stem cell base therapy is to extensive
It spreads the disease, as the treatment and healing of diabetes, neurodegeneration or cardiovascular disease have great influence.Meanwhile it is the first dry thin
The internal transplantation experiments of born of the same parents' tracheae implantation achieve successfully (Macchiarini P.; Jungebluth P.; Go T.;
Asnaghi M. A.; Rees L. E.; Cogan T. A.; Dodson A.; Martorell J.; Bellini S.;
Parnigotto P. P. Clinical transplantation of a tissue-engineered airway[J].
The Lancet, 2008,372,2023-2030.), and stem cell can be tracked in vivo, enable us to monitor its life
Object distribution and its existence in histoorgan, and understand stem cell and the cell of receptor after transplanting interact and mode.
In recent years studies have shown that fluorescent nano particle field of biomedicine application in the past few decades in cause
Great research interest (Michalet, X.; Pinaud, F. F.; Bentolila, L. A.; Tsay, J. M.;
Doose, S.;Li, J. J.; Sundaresan, G.; Wu, A. M.; Gambhir, S. S.; Weiss, S.
Quantum Dots for Live Cells in Vivo Imaging, and Diagnostics. Science, 2005,
307, 538−544. Wang, K.; He, X.; Yang, X.; Shi, H. Functionalized Silica
Nanoparticles: A Platform for Fluore-scence Imaging at the Cell and Small
Animal Levels. Acc. Chem. Res. 2013, 46, 1367−1376. Zhou, J.; Liu, Z.; Li, F.
Upconversion Nanophosphors for Small-animal Imaging. Chem. Soc. Rev. 2012,
41, 1323−1349.).Fluorescent nano particle is such as: quantum dot, upper conversion nano particle and the silica nanometer based on dyestuff
Particle can pass through fluorescence imaging real-time visual or monitoring vivo biodistribution.Wherein, the nano SiO 2 particle based on dyestuff is steady
Qualitative height, is easy to surface modification, has good biocompatibility, has obtained systematic research (Montalti, M.;
Prodi, L.; Rampazzo, E.; Zaccheroni, N. Dye[1]Doped Silica Nanoparticles as
Luminescent Organized Systems for Nanomedicine. Chem. Soc. Rev. 2014, 43,
4243−4268.).However, light drift often occurs after traditional dyestuff mixes in the nano SiO 2 particle of high concentration
Quenching caused by white and aggregation, to hinder its further applying in bio-imaging.Due to aggregation-induced emission light source
The discovery of AIE, different AIE, which is embedded into nano SiO 2 particle by way of physical package or covalent coupling, carries out height
Bio-imaging (Luo, the J. of effect; Xie, Z.; Lam, J. W. Y.; Cheng, L.; Chen, H.; Qiu, C.;
Kwok, H. S.; Zhan, X.; Liu, Y.; Zhu, D.; Tang, B. Z. Aggregation[1]Induced
Emission of 1-Methyl-1,2,3,4,5-pentaphenylsilole. Chem. Commun. 2001, 1740-
1741. Ding, D.; Li, K.; Liu, B.; Tang, B. Z. Bioprobes Based on AIE
Fluorogens. Acc. Chem. Res. 2013, 46, 2441−2453. Hong, Y.; Lam, J. W. Y.;
Tang, B. Z. Aggregation-Induced
Emission: Phenomenon, Mechanism and Applications. Chem.Commun. 2009, 4332
−4353.).Due to Gd3+It is a kind of paramagnetic material, with superparamagnetic material phase for common MR molecular imaging signal component
Than Gd3+There is lower light absorptive to fluorescent material, be not easy to cause fluorescent quenching effect, can be used as bimodal image probe
MR imaging signal component.
Based on the above research background, the spy of the fluorescence magnetic particle that synthesizes herein with quantum dot and magnetic particle
Matter is wrapped up, good biocompatibility by silicon shell, and can highly efficient labeling human adipose mesenchymal stem cells, fluorescence imaging can be passed through
With the labeled situation of the external dynamic monitoring mescenchymal stem cell of magnetic resonance imaging double mode, therefore can be used as it is a kind of it is safe,
Ideal stem cell tracer, the experimental study applied to stem cell.
Summary of the invention
The technical problems to be solved by the present invention are: how to prepare a kind of bioprobe of bimodal imaging, and have compared with
Good biocompatibility and lower toxicity can mark mescenchymal stem cell as probe, and be able to achieve fluorescence and nuclear-magnetism
Bimodal imaging.
The technical scheme adopted by the invention is that: a kind of method of bimodal nano-probe preparation, in accordance with the following steps into
Row
Step 1: mesoporous silicon oxide is sintered into fine silica end, by Gd (NO3)3·6H2O dissolves in deionized water
Form Gd (NO3)3Solution, then by Gd (NO3)3Solution is added dropwise in fine silica end, and dropwise addition process is stirred
It mixes, then drying and calcination, obtains Gd@SiO2Compound；
Step 2: aggregation-induced emission AIE fluorescent dye is dissolved in formation AIE solution in acetonitrile solution, then by AIE solution
It is added drop-wise to Gd@SiO dropwise2In compound, it is dried after being uniformly mixed, is then added to polyvinylpyrrolidone and second
In the mixed solution of glycol, the ammonium hydroxide that mass percent concentration is 25% is added after mixing evenly, then adds ethyl alcohol and just
Silester is finally cleaned to remove the unbonded AIE dyestuff in surface with acetonitrile, sediment is dried, is obtained
AIE-Gd@SiO2Nano-complex is bimodal nano-probe.
In step 1, the amount of mesoporous silicon oxide is 0.1-1 grams, and the amount at the fine silica end formed after calcining is
0.06-0.6 grams, Gd (NO3)3·6H2The quality of O is 0.06-0.6 grams, Gd (NO3)3The concentration of solution is 0.4-4 mol/L.
In step 2,0.02-0.2 grams of aggregation-induced emission AIE fluorescent dye is dissolved in 300-2000 microlitres of second
AIE solution is formed in nitrile solution, and AIE solution is then added drop-wise to 0.08-0.7 grams of Gd@SiO dropwise2In compound, then it is added
To in the mixed solution of 0.25-2.5 grams of polyvinylpyrrolidone and 25ml ethylene glycol, add 2.5-25ml's after mixing evenly
The ammonium hydroxide that mass percent concentration is 25%, then adds 1.25-12.5ml ethyl alcohol and 25-250ul ethyl orthosilicate.
The AIE-Gd SiO for being first 1mg/ml with deionized water ultrasound arrangement concentration2Nano-complex aqueous solution, later
Nano-complex aqueous solution is diluted to different concentration with culture medium, therewith by the AIE-Gd@SiO of various concentration2Nanometer is multiple
It closes object to be incubated in the incubator with mescenchymal stem cell respectively for 24 hours, then is cleaned to remove three times with phosphate buffer solution and do not gulped down
The AIE-Gd@SiO bitten2Nano-complex is then added the culture medium containing thiazolyl blue MTT solution and continues to cultivate 4h, is added two
First sulfoxide solution low-speed oscillation 10min, the light absorption value of each culture hole is surveyed with microplate reader under the wavelength of 490nm；Cell is increased
It grows experiment, is incubated for after cleaning for 24 hours and continues to cultivate 5d, method same as above surveys absorbance, surveys cytotoxicity with this.
By the AIE-Gd@SiO of various concentration2Nano material is same as stem cell be incubated for for 24 hours after, clean, it is fixed, be placed in sharp
Its fluorescence imaging is observed under light Laser Scanning Confocal Microscope；For NMR imaging, the nano-complex of cell and various concentration is incubated for
After for 24 hours, recombination pancreatin digestion centrifugation rear overhang in the PCR pipe that phosphate buffer solution and agarose solution mix, by nuclear-magnetism at
As strong and weak to observe magnetic signal of the stem cell of nano combined substance markers in agarose solution.
The AIE-Gd@SiO of various concentration2Nano material refer to 5 mcg/mls, 10 mcg/mls, 20 mcg/mls,
40 mcg/mls, 80 mcg/mls, phosphate buffer solution are 50-500 microlitres, and the amount of agarose is 0.05g, and mass fraction is
1%, volume is 50-500 microlitres.
The beneficial effects of the present invention are: the method for the present invention prepares AIE-Gd@SiO2Nano-complex has good biology
Compatibility and hypotoxicity can be used as nano-probe label mescenchymal stem cell, while fluorescence and nuclear-magnetism bimodal may be implemented
Imaging, the experimental study applied to stem cell.
Detailed description of the invention
Fig. 1 is AIE-Gd@SiO2Transmission electron microscope (TEM) figure of compound；
Fig. 2 is AIE-Gd@SiO2The cell activity figure of the mescenchymal stem cell of compound substance markers, wherein abscissa AIE-Gd
SiO2Concentration (ul/ml), ordinate are cell activity (%)；
Fig. 3 is cell Proliferation figure, and wherein abscissa is AIE-Gd@SiO2Concentration (ul/ml), ordinate are cell Proliferation (%),
Time is 5 days；
Fig. 4 is AIE-Gd@SiO2The laser confocal microscope image of nano combined substance markers mescenchymal stem cell；
Fig. 5 is AIE-Gd@SiO2The NMR imaging of nano combined substance markers mescenchymal stem cell.
Specific embodiment
The preparation of bimodal nano-probe and its to the label of mescenchymal stem cell
0.1-1 grams of mesoporous silicon oxide is first carried out to high-temperature calcination in Muffle furnace into fine powder, then compound 30% gadolinium.This mistake
Journey is by by 0.06-0.6 grams of gadolinium nitrate hexahydrate [Gd (NO3)3·6H2O] solid dissolution be configured to solution in deionized water,
Then the solution is added dropwise in meso-porous titanium dioxide Si powder, is dried after mixing and high-temperature calcination, obtain Gd
SiO2Compound；
Step 2: taking 0.02-0.2 grams of aggregation-induced emission (AIE) fluorescent dye ultrasonic dissolution molten in 300-2000 microlitres of acetonitrile
In liquid, by 0.08-0.7 grams of Gd@SiO of synthesis2Composite material is put into surface plate is added dropwise AIE solution dropwise, is stirred for mixing
It is dried after closing uniformly.Then 0.05-0.5 grams of compound after taking drying, while 0.25-2.5 grams of polyvinyl pyrrole is added
12h is mixed in alkanone and 25ml ethylene glycol, adds 2.5-25ml(25%) ammonium hydroxide ultrasound 30min after 1.25- is added
12.5ml ethyl alcohol and 25-250ul ethyl orthosilicate ultrasound 1h finally carry out eccentric cleaning with acetonitrile and keep supernatant colourless, dry,
That is AIE-Gd@SiO2Nano-complex.For the dispersibility with the nano material of synthesis, first by compound good nanometer material
Material be dispersed in solution made of polyvinylpyrrolidone and deionized water, add 0.01-0.15 grams of polyethylene glycol ultrasound observation its
Dispersibility.
Utilize the AIE-Gd@SiO of preparation2Compound is as nano-probe labeled stem cells, it is characterised in that: will be different dense
Spend the AIE-Gd@SiO of 5 mcg/mls, 10 mcg/mls, 20 mcg/mls, 40 mcg/mls, 80 mcg/mls2Nanometer
Compound and mescenchymal stem cell are incubated for for 24 hours, then with 5-15 milliliters of phosphate buffer solutions clean three times with removing not by
The nano particle of phagocytosis is then added the culture medium containing 0.25g thiazolyl blue solution and continues to cultivate 4h, sucks culture medium, be added
150-5300ul dimethyl sulfoxide solution low-speed oscillation 10min, the extinction of each culture hole is surveyed with microplate reader under the wavelength of 490nm
Value；It for cell proliferation experiment, is incubated for after cleaning for 24 hours and continues to cultivate 5d, method same as above surveys absorbance, thin to survey with this
Cellular toxicity.
Fluorescence is carried out using the labeled stem cells of preparation and nuclear-magnetism bimodal is imaged, it is characterised in that: various concentration 5 is micro-
The AIE-Gd@SiO of grams per milliliter, 10 mcg/mls, 20 mcg/mls, 40 mcg/mls, 80 mcg/mls2Nano material
It is same as stem cell be incubated for for 24 hours after, clean, it is fixed, be placed under laser confocal microscope and observe its fluorescence imaging；For nuclear-magnetism
Imaging, after the nano-complex of cell and various concentration is incubated for for 24 hours, recombination pancreatin digestion centrifugation rear overhang is in 50-500 microlitres of phosphorus
In acid buffering solution and the PCR pipe of 0.05g agarose solution mixing, the dry of nano combined substance markers is observed by NMR imaging
Magnetic signal of the cell in agarose solution is strong and weak.The experimental results showed that the stem cell of nano-probe label can carry out fluorescence
It is imaged with nuclear-magnetism bimodal.
Claims (7)
1. a kind of method of bimodal nano-probe preparation, it is characterised in that: carry out in accordance with the following steps
Step 1: mesoporous silicon oxide is sintered into fine silica end, by Gd (NO3)3·6H2O dissolves in deionized water
Form Gd (NO3)3Solution, then by Gd (NO3)3Solution is added dropwise in fine silica end, and dropwise addition process is stirred
It mixes, then drying and calcination, obtains Gd@SiO2Compound；
Step 2: aggregation-induced emission AIE fluorescent dye is dissolved in formation AIE solution in acetonitrile solution, then by AIE solution
It is added drop-wise to Gd@SiO dropwise2In compound, it is dried after being uniformly mixed, is then added to polyvinylpyrrolidone and second
In the mixed solution of glycol, the ammonium hydroxide that mass percent concentration is 25% is added after mixing evenly, then adds ethyl alcohol and just
Silester is finally cleaned to remove the unbonded AIE dyestuff in surface with acetonitrile, sediment is dried, is obtained
AIE-Gd@SiO2Nano-complex is bimodal nano-probe.
2. the method for bimodal nano-probe preparation according to claim 1, it is characterised in that: in step 1, mesoporous two
The amount of silica is 0.1-1 grams, and the amount at the fine silica end formed after calcining is 0.06-0.6 grams, Gd (NO3)3·6H2O
Quality be 0.06-0.6 grams, Gd (NO3)3The concentration of solution is 0.4-4 mol/L.
3. the method for bimodal nano-probe preparation according to claim 1, it is characterised in that: in step 2, by 0.02-
0.2 gram of aggregation-induced emission AIE fluorescent dye is dissolved in formation AIE solution in 300-2000 microlitres of acetonitrile solution, then
AIE solution is added drop-wise to 0.08-0.7 grams of Gd@SiO dropwise2In compound, it is then added to 0.25-2.5 grams of polyvinylpyrrolidine
In the mixed solution of ketone and 25ml ethylene glycol, the ammonia that the mass percent concentration of 2.5-25ml is 25% is added after mixing evenly
Then water adds 1.25-12.5ml ethyl alcohol and 25-250ul ethyl orthosilicate.
4. utilizing bimodal nano-probe labeled stem cells, it is characterised in that: be with deionized water ultrasound arrangement concentration first
The AIE-Gd@SiO of 1mg/ml2Nano-complex aqueous solution is diluted to difference with culture medium later by nano-complex aqueous solution
Concentration, therewith by the AIE-Gd@SiO of various concentration2Nano-complex is incubated for mescenchymal stem cell in the incubator respectively
For 24 hours, the AIE-Gd@SiO for removing do not swallowed three times is then cleaned with phosphate buffer solution2Nano-complex is then added and contains
There is the culture medium of thiazolyl blue MTT solution to continue to cultivate 4h, dimethyl sulfoxide solution low-speed oscillation 10min is added, is existed with microplate reader
The light absorption value of each culture hole is surveyed under the wavelength of 490nm；For cell proliferation experiment, it is incubated for and continues to cultivate 5d after cleaning for 24 hours, ibid
The method surveys absorbance, surveys cytotoxicity with this.
5. according to claim 4 utilize bimodal nano-probe labeled stem cells, it is characterised in that: various concentration
AIE-Gd@SiO2Nano-complex is 5 mcg/mls, 10 mcg/mls, 20 mcg/mls, 40 mcg/mls, 80 respectively
Mcg/ml, the amount of thiazolyl blue are 0.25g, concentration 5mg/ml；The amount of phosphate buffer is 5-15 milliliters, pH 7.4；Two
The amount of first sulfoxide is 150-5300ul.
6. bimodal nano-probe labeled stem cells carry out fluorescence and the imaging of nuclear-magnetism bimodal, it is characterised in that: by various concentration
AIE-Gd@SiO2Nano material is same as stem cell be incubated for for 24 hours after, clean, it is fixed, be placed under laser confocal microscope and see
Examine its fluorescence imaging；For NMR imaging, after the nano-complex of cell and various concentration is incubated for for 24 hours, recombination pancreatin digestion
Rear overhang is centrifuged in the PCR pipe that phosphate buffer solution and agarose solution mix, nano-complex is observed by NMR imaging
Magnetic signal of the stem cell of label in agarose solution is strong and weak.
7. bimodal nano-probe labeled stem cells according to claim 6 carry out fluorescence and the imaging of nuclear-magnetism bimodal,
It is characterized in that: the AIE-Gd@SiO of various concentration2Nano material refer to 5 mcg/mls, 10 mcg/mls, 20 mcg/mls,
40 mcg/mls, 80 mcg/mls, phosphate buffer solution are 50-500 microlitres, and the amount of agarose is 0.05g, and mass fraction is
1%, volume is 50-500 microlitres.
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