CN107478618A - A kind of method of the intracellular ROS detections based on fluorescence gold nanoclusters - Google Patents
A kind of method of the intracellular ROS detections based on fluorescence gold nanoclusters Download PDFInfo
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Abstract
The present invention provides a kind of preparation method of the arginic fluorescence gold nanoclusters of surface modification oligomerization and the fluorescence gold nanoclusters, and its active oxygen detection method based on the fluorescence gold nanoclusters and the kit for including the fluorescence gold nanoclusters, and the application of the fluorescence nano cluster and the combination product.The fluorogold nano-cluster has powerful anti-light Bleachability, anti-light glitter and good biocompatibility.And the active oxygen detection method has that method is simple, reaction condition is gentle, i.e. the characteristic of normal temperature and pressure, aqueous phase and pH near neutrals, be advantageous to the biologic applications of the fluorescence gold nanoclusters.
Description
Technical field
The present invention relates to a kind of fluorescence gold nanoclusters, more particularly to a kind of arginic fluorescence gold nano of surface modification oligomerization
The preparation method of cluster and the fluorescence gold nanoclusters, and its active oxygen (ROS) detection method based on the fluorescence gold nano and comprising
The kit of the fluorescence gold nanoclusters, and the application of the fluorescence nano cluster and the kit.
Background technology
In order to observe intracellular ROS situation, researcher designs and synthesizes the inspection that organic fluorescence probe is used for ROS
Survey.There are some researches show organic fluorescence probe has high selectivity to specific ROS, and in the physiological mechanism for studying specific ROS
Method has great potential.However, the photobleaching of these organic fluorescence probes, the limitation such as bio-toxicity and spontaneous oxidative
Property hinders its extensive and prolonged application in cell.
ROS is caused important signaling molecule in metabolic process, and it includes superoxides (O2 -), singlet oxygen (O2 1),
Hydrogen peroxide (H2O2), hydroxyl free radical (OH), hypochlorite (ClO-), Peroxynitrite (ONOO-) etc..Active oxygen with very
More serious human diseases such as cancer, angiocardiopathy, senile dementia and nerve degenerative diseases etc. are relevant.Research table recently
Bright ROS plays the part of pivotal player in various physiological functions are adjusted as second messenger, therefore ROS has attracted many chemistry, biology
The researcher in the field such as and medical science is studied it.Also, in various active oxygens, OH, ClO-And ONOO-It is considered as
It is high activity oxygen (hROS), because their Strong oxdiative characteristic, can be with the nucleic acid in direct oxidation living cells, albumen
Matter, lipid etc., cause potential serious infringement.Therefore, detections of the hROS in cellular environment be also very important, and it can be with
Us are helped to more fully understand physiological actions of the hROS in living cells.
The content of the invention
Therefore, based on above-mentioned prior art the defects of, it is an object of the invention to provide a kind of surface modification oligomerization arginine
Fluorescence gold nanoclusters and the fluorescence gold nanoclusters preparation method, and its active oxygen (ROS) based on the fluorescence gold nanoclusters
Detection method and the kit for including the fluorescence gold nanoclusters, and the application of the fluorescence nano cluster and the kit.
For foregoing invention purpose, the first aspect of the present invention provides a kind of fluorescence gold nanoclusters, the fluorescence Jenner
The surface modification of rice cluster has oligomerization arginine.The oligomerization arginine is preferably the small peptide of eight to two ten arginine compositions.Most
Preferably nine poly arginines.
The second aspect of the present invention provides a kind of preparation method, and it is used for the fluorescence Jenner for preparing first aspect present invention
Rice cluster, wherein, the preparation method includes：Gold chloride, oligomerization arginine peptide and protective agent are mixed to get mixed liquor；By described in
Mixed liquor heated at constant temperature synthesizes the fluorescence gold nanoclusters.
Preparation method according to a second aspect of the present invention, wherein, the protective agent is selected from glutathione, ascorbic acid and half
One or more in cystine.
Preparation method according to a second aspect of the present invention, wherein：Mole of the gold chloride and the oligomerization arginine peptide
Than for 0.1~10：1, preferably 0.2~2.5：1, most preferably 2.4:1.And/or the protective agent with oligomerization is arginic rubs
You are than being 0~10:1, most preferably 2:1.
Preparation method according to a second aspect of the present invention, wherein：The heated at constant temperature is 40~120 DEG C, is preferably
50~100 DEG C, most preferably 70 DEG C.And/or the heated at constant temperature time is 12~48h, preferably 18~30h, is most preferably
24h。
The third aspect of the present invention provides a kind of active oxygen detection method, its fluorogold based on first aspect present invention
Nano-cluster or the fluorescence gold nanoclusters for using the method for second aspect of the present invention to prepare, wherein, the detection method includes：
(1) the fluorescence gold nanoclusters are added into sample to be tested and are reacted；Preferably,
The reaction temperature be 20~40 DEG C, more preferably 20~37 DEG C, most preferably 25 DEG C or 37 DEG C,
The reaction time is 10~60min, more preferably 10~30min, most preferably 15min or 20min,
The concentration of the fluorescence gold nanoclusters is 100~400 μ g/ml, more preferably 200~400 μ g/ml, is more entered
One step is preferably 200~300 μ g/ml, and/or
The volume of the fluorescence gold nanoclusters is 100~300 μ l, more preferably 150~250 μ l, further excellent
Elect 160~200 μ l as；
(2) fluorescence of the sample to be tested is measured to detect the presence of the active oxygen or concentration；
Preferably, the sample to be tested is solution or cell, and/or
Preferably, the active oxygen be selected from superoxides, singlet oxygen, hydrogen peroxide, hydroxyl free radical, hypochlorite and
One or more in Peroxynitrite.
Active oxygen detection method according to a third aspect of the present invention, wherein, when the sample to be tested is solution, step
(2) presence of the detection active oxygen or concentration are carried out by calibration curve method in.Preferably, the step (2) includes drawing
Standard curve, draw the standard curve and comprise the following steps：(a) described in being added into the active oxygen solutions of various concentrations
Fluorescence gold nanoclusters simultaneously react, and (b) detects the fluorescence intensity of the active oxygen solutions, according to the fluorescence intensity and the activity
The concentration of oxygen solution draws standard curve.It is highly preferred that the concentration of the active oxygen solutions be respectively 0nM, 40nM, 100nM,
200nM, 400nM, 1 μM, 4 μM, 20 μM, 40 μM, 60 μM, 80 μM and 250 μM.
Active oxygen detection method according to a third aspect of the present invention, wherein, when the sample to be tested is cell, step
(1) reaction in is incubation.Preferably, after step (1), the detection method also include washing away do not enter into it is described
The fluorescence gold nanoclusters of cell.
In one embodiment, above-mentioned active oxygen detection method can include：
(1) by the fluorescence gold nanoclusters and cell incubation；
(2) the fluorescence gold nanoclusters for not entering into the cell are washed away；
(3) fluorescence of the cell is measured to detect the presence of the active oxygen or concentration.
Wherein, the fluorescence of the measurement cell can use ELIASA measurement optical density (OD) value or streaming in step (3)
Cell instrument measures fluorescence intensity.
The fourth aspect of the present invention provides a kind of kit, and the kit includes the fluorescence Jenner of first aspect present invention
Rice cluster or the fluorescence gold nanoclusters prepared using the preparation method of second aspect of the present invention.
The fifth aspect of the present invention provides a kind of application, the fluorescence gold nanoclusters of the invention first aspect that bases on practicality,
It is prepared by the fluorescence gold nanoclusters or the kit of fourth aspect present invention prepared using the preparation method of second aspect of the present invention
For detecting the application in the product selected from one or more of illness：Cancer, angiocardiopathy, senile dementia and nerve
Degenerative disease.
The sixth aspect of the present invention provides a kind of method for detecting and being selected from one or more of illness：Cancer, painstaking effort
Pipe disease, senile dementia and nerve degenerative diseases, the active oxygen detection method that methods described passes through third aspect present invention
Carry out.
The seventh aspect of the present invention provides a kind of fluorescence gold nano for being used for detection and being selected from one or more of illness
Cluster：Cancer, angiocardiopathy, senile dementia and nerve degenerative diseases, the fluorescence gold nanoclusters are first party of the present invention
The fluorescence gold nanoclusters in face or the fluorescence gold nanoclusters prepared using the preparation method of second aspect of the present invention.
Specifically, should the invention provides the fluorescence gold nanoclusters of utilization one-step synthesis method oligomerization arginine modification and utilization
Fluorescence gold nanoclusters detect the ROS in cell as nano-sensor.The fluorescence gold nanoclusters are as a kind of new spy
Pin is different from traditional organic fluorescence molecular probe, and it has powerful anti-light Bleachability, anti-light glitter and good biofacies
Capacitive etc..The fluorescence gold nanoclusters can pass through cell membrane to enter cell and be sent under excited by visible light stronger red glimmering
Light.When producing excessive ROS into the cell, the fluorescence of Jenner is quenched and the degree of fluorescent quenching and intracellular ROS amount are
Wired sexual intercourse.Therefore monitor and detect in real time the ROS in cell using described this characteristic of fluorescence gold nanoclusters.This hair
The bright detection for intracellular ROS opens a kind of promising new method.
Fluorescence gold nanoclusters of the present invention are prepared by one-step method, and this substantially reduces generated time, are improved
Combined coefficient.Compared with the organic fluorescence molecule used in commercial kits, described fluorogold nano-cluster has more preferable
Optical property and biocompatibility, stronger anti-light Bleachability and lower bio-toxicity, be advantageous to detect for a long time.The present invention
The active oxygen detection method based on the fluorescence gold nanoclusters provided has that method is simple, reaction condition is gentle, i.e., normal temperature is normal
The characteristic of pressure, aqueous phase and pH near neutrals, be advantageous to the biologic applications of the fluorescence gold nanoclusters.And the fluorescence nano cluster table
Face is modified with cell-penetrating peptide, and this preferably helps the fluorescence nano cluster to enter cell, so as to realize intracellular ROS detection.
Brief description of the drawings
Hereinafter, embodiment of the present invention is described in detail with reference to accompanying drawing, wherein：
Fig. 1 shows fluorescence gold nanoclusters (AuNCs) synthesis and application schematic diagram, wherein HAuCl4As chlorine gold
Acid, GSH are glutathione, and AuNCs is fluorescence gold nanoclusters；Hv is illumination, and hROS is high activity oxygen, Em：
620nm is the fluorescence for launching 620nm wavelength；
Fig. 2 shows the ultraviolet-visible absorption spectroscopy, fluorescence spectrum and the hair when exciting light is 430nm of the AuNCs
Penetrate spectrum, and the solution of the AuNCs and solution fluorescence figure；
Fig. 3 shows the fluorescence spectra that the AuNCs responds for the OH of various concentrations；
Fig. 4 shows that the AuNCs is used for the standard curve that ROS is detected in solution；
Fig. 5 shows the selectivity experiment that the AuNCs detects to ROS；
Fig. 6 shows the AuNCs toxicity tests result detected using CCK-8 kits；
Fig. 7 shows cellular morphology figure when detecting the AuNCs toxicity；
Fig. 8 shows the fluorogram of HUVEC AuNCs different times under light illumination into the cell；
Fig. 9 shows HUVEC AuNCs into the cell fluorescence curve figure；
Figure 10, which is shown, utilizes the AuNCs and the intracellular ROS measured by commercial kits；
Figure 11 shows the flow cytometry results that intracellular ROS is detected using the AuNCs；
Figure 12 shows the flow cytometry results that intracellular ROS is detected using commercial kits；
Description of reference numerals：
1st, the AuNCs solution figure；2nd, the fluorogram of the AuNCs solution；3 and the cellular morphology figure of 5, control group；4 Hes
6th, the cellular morphology figure added after the AuNCs.
Embodiment
The present invention is further illustrated below by specific embodiment, it should be understood, however, that, these embodiments are only
It is used for specifically describing in more detail, and is not to be construed as limiting the present invention in any form.
In following examples, in addition to the test material, condition and the operating method that particularly point out, used in embodiment
Many materials and operating method are well known in the art.Therefore, it will be apparent to those skilled in the art that within a context, if not special
Do not mentionlet alone bright, material therefor of the present invention and operating method are well known in the art.
The reagent and instrument used in following examples is as follows：
Reagent：
Gold chloride, glutathione, potassium peroxide, hydrogen peroxide, copper chloride, Peroxynitrite iron, purchased from Sigma；
Oligomerization arginine synthesizes purchased from Shanghai gill biochemical corp；
CCK-8 kits, purchased from DOJINDO；
ROS commercial kits, purchased from eBioscience.
Instrument：
ELIASA, purchased from Perkin Elmer companies of the U.S., model Enspire；
Ultraviolet specrophotometer, purchased from Shimadzu, Japan, model UV-2450；
XRF, purchased from Shimadzu, Japan, model RF-5301PC；
Flow cytometer, purchased from U.S. company BD, model BD LSRFortessa SORP.
Concentration unit mM is mmol/L, and μM as μm ol/L, nM is nmol/L.
Embodiment 1：Synthesize fluorescence gold nanoclusters
The present embodiment is used to illustrate fluorescence gold nanoclusters provided by the invention.
Described fluorescence gold nanoclusters preparation method such as Fig. 1, it is specially：
Mix the gold chloride (HAuCl that 300 μ l concentration are 120mM4), 10ml concentration be 1.5mM part (Ligand, i.e.,
Oligomerization arginine peptide) and 10ml concentration be 3mM glutathione (GSH) obtain mixed liquor；By described 70 DEG C of heating 24 of mixed liquor
Hour synthesizes the fluorescence gold nanoclusters (AuNCs).
Fluorescence gold nanoclusters solution such as Fig. 2 references 1 synthesized by the embodiment, color is chartreuse, the fluorescence
The fluorescence color of gold nanoclusters solution such as Fig. 2 references 2, color is Chinese red.Using described in fluorescence spectrophotometer measurement
Absorption spectrum, excitation spectrum and emission spectrum such as Fig. 2 under 430nm wavelength of fluorescence gold nanoclusters solution.
Test example 1：The sensitivity experiment that ROS is detected in solution
The AuNCs synthesized using embodiment 1 carries out the detection of ROS in solution.
The detection sensitivity is tested：
ROS in the solution is as the OH prepared by Fenton's reaction.
To various concentrations (0nM, 40nM, 100nM, 200nM, 400nM, 1 μM, 4 μM, 20 μM, 40 μM, 60 μM, 80 μM and
250 μM) the OH aqueous solution in be separately added into 180 μ l concentration be 300 μ g/ml AuNCs and react 15min at 25 DEG C；Adopt
Fluorescence spectrum is measured with ELIASA.
Part fluorescence spectrum such as Fig. 3 of the detection sensitivity, illustrate as the increase of ROS concentration, fluorescence can subtract step by step
It is weak.It was found from test result, the AuNCs as nano-sensor be used for solution in detection ROS have good sensitivity and
Test limit, and minimum test limit can reach 100nM.
Test example 2：Draw the standard curve of ROS in detection solution
The AuNCs synthesized using embodiment 1 draw the standard curve of ROS in detection solution.
Specific method includes：
(a) to various concentrations (0nM, 40nM, 100nM, 200nM, 400nM, 1 μM, 4 μM, 20 μM, 40 μM, 60 μM, 80 μM
With 250 μM) the OH aqueous solution in be separately added into 180 μ l concentration be 300 μ g/ml AuNCs and react 20min at 25 DEG C；
(b) fluorescence intensity of the OH aqueous solution is detected, is painted according to the concentration of the fluorescence intensity and the active oxygen solutions
Standard curve processed.
The standard curve such as Fig. 4, abscissa are OH concentration, and ordinate is (F0-F)/F0Result of calculation, wherein F
For the fluorescence intensity (FL intensity) of the various concentrations solution, F0For FL intensity when OH concentration is 0 in solution.
Test example 3：Detect the selectivity experiment of ROS in solution
The AuNCs synthesized using embodiment 1 carries out the detection of ROS in solution.
The selectivity, which is tested, is specially：
To solution to be measured (100 μM of ClO is included respectively-, 100 μM of ONOO-, 100 μM OH, 1mM H2O2、1mM
O2 -And typically occur in other materials in biological matrix, i.e. 1mM Ca2+, 1mM Zn2+, 1mM Fe2+, 1mM paddy
Propylhomoserin (Glutamic acid), 1mM glycine (Glycine), 1mM glucose (Glucose) and 1mM ox blood are pure
Albumen (BSA)) in be separately added into 180 μ l concentration be 300 μ g/ml AuNCs and react 20min at 25 DEG C；Using ELIASA
Measure the solution fluorescence intensity.
Test result such as Fig. 5, it was found from test result, the AuNCs can detect ClO in solution-、ONOO-And OH.
Test example 4：Cytotoxicity and cellular morphology observation experiment
The AuNCs toxicity of the synthesis of embodiment 1 is detected in CCK-8 kits using HUVEC cells and A375 cells.
Cytotoxicity experiment is specially：
The AuNCs is subjected to gradient dilution, ultimate density is respectively 0 μ g/mL, 8 μ g/mL, 16 μ g/mL, 32 μ after dilution
G/mL, 64 μ g/mL, 160 μ g/mL and 320 μ g/mL；The AuNCs aqueous solution of 10 μ l various concentrations is separately added into certain amount
Cell in and at 37 DEG C be incubated 24h；Add CCK-8 reagents；Its OD value at 450nm is measured with ELIASA.
Cellular morphology observation experiment is specially：
The AuNCs aqueous solution that 10 μ l concentration are 160 μ g/mL is separately added into the cell；It is incubated at 37 DEG C
24 hours, add whether AuNCs has an impact to the form of cell using micro- sem observation.
Cytotoxicity experiment result such as Fig. 6, from test result, when AuNCs concentration of aqueous solution is less than 160 μ g/ml, carefully
The survival rate of born of the same parents is up to more than 85%.Cellular morphology observation experiment such as Fig. 7, compared with control group, that is, it is not added with the thin of the AuNCs aqueous solution
Born of the same parents, the form of cell do not have significant change.In summary, the cytotoxicity of the fluorescence gold nanoclusters is small, there is preferable life
Thing compatibility.
Test example 5：Intracellular ROS imagings
The 20 μ L concentration that embodiment 1 is synthesized are that 320 μ g/mL AuNCs and HUVEC cell is incubated 3 hours at 37 DEG C；
Then the AuNCs for not entering into cell is washed away；Different ROS (H are added into the cell to AuNCs be present2O2Or HClO)；Simultaneously
Observed under fluorescence co-focusing and record a data every 3min.
Testing result such as Fig. 8 and Fig. 9, wherein Fig. 9 abscissa are observing time, ordinate F/F0Result of calculation, F
For the fluorescence intensity (FL intensity) of the various concentrations solution, F0For FL intensity when ROS concentration is 0 in solution.Tied by test
Fruit understands that fluorescence signals of the AuNCs under continuous illumination in cell is without significant change.
Test example 6：Detect intracellular ROS
The 20 μ L concentration that embodiment 1 is synthesized first be 320 μ g/mL AuNCs and ROS commercial kits in probe with
Cell is incubated 20min at 37 DEG C；Wash away the AuNCs and probe for not entering into cell；Various concentrations are added into the cell
The H of (0,0.1,0.2,0.4,1 and 2mM)2O2, then go to survey the fluorescence intensity of each cell and statistical using flow cytometer
Analysis.
Figure 10 is the H that the AuNCs and commercial kits measure various concentrations2O2The test result of solution, wherein, Figure 10
Abscissa is H2O2The concentration of solution, ordinate are (F0-F)/F0Result of calculation, wherein F be the various concentrations solution it is glimmering
Luminous intensity (FL intensity), F0For H in solution2O2FL intensity when concentration is 0.Figure 11 is to detect intracellular H by the AuNCs2O2
Flow cytometry results；Figure 12 is to detect intracellular H by the commercial kits2O2Flow cytometry results.As a result table
Bright, the testing result detected by AuNCs is consistent with the testing result trend detected with the commercial kits, and without notable
Difference.
Although here, a certain degree of description has been carried out to the present invention, it will be apparent that, do not depart from the present invention spirit and
Under conditions of scope, one of ordinary skill in the art can carry out the appropriate change of each condition.It is understood that the present invention is unlimited
Summarize in the embodiment and instantiation, its right are attributed to the scope of claim, and including each factor etc.
With replacement.
Claims (10)
1. a kind of fluorescence gold nanoclusters, it is characterised in that the surface modification of the fluorescence gold nanoclusters has oligomerization arginine；It is described
Oligomerization arginine is preferably the small peptide of eight to two ten arginine compositions；Most preferably nine poly arginines.
2. the preparation method of the fluorescence gold nanoclusters described in claim 1, it is characterised in that methods described includes：
Gold chloride, oligomerization arginine peptide and protective agent are mixed to get mixed liquor；
The mixed liquor heated at constant temperature is synthesized into the fluorescence gold nanoclusters.
3. the preparation method of fluorescence gold nanoclusters according to claim 2, it is characterised in that the protective agent is selected from paddy Guang
One or more in sweet peptide, ascorbic acid and cysteine.
4. the preparation method of the fluorescence gold nanoclusters according to Claims 2 or 3, it is characterised in that：
The mol ratio 0.1~10 of the gold chloride and the oligomerization arginine peptide:1, preferably 0.2~2.5:1, be most preferably
2.4:1；And/or
Protective agent is 0~10 with the arginic mol ratio of oligomerization:1, most preferably 2:1.
5. the preparation method of the fluorescence gold nanoclusters according to any one of claim 2 to 4, it is characterised in that：
The temperature for stating heated at constant temperature is 40~120 DEG C, preferably 50~100 DEG C, most preferably 70 DEG C, and/or
12~48h of time of the heated at constant temperature, most preferably preferably 18~30h, 24h.
6. a kind of active oxygen detection method, the detection method is based on fluorescence gold nanoclusters described in claim 1 or using right
It is required that fluorescence gold nanoclusters prepared by method any one of 2 to 5, it is characterised in that the detection method includes：
(1) the fluorescence gold nanoclusters are added into sample to be tested and are reacted；Preferably,
The reaction temperature be 20~40 DEG C, more preferably 20~37 DEG C, most preferably 25 DEG C or 37 DEG C,
The reaction time is 10~60min, more preferably 10~30min, most preferably 15min or 20min,
The concentration of the fluorescence gold nanoclusters is 100~400 μ g/ml, more preferably 200~400 μ g/ml, further
Preferably 200~300 μ g/ml, and/or
The volume of the fluorescence gold nanoclusters is 100~300 μ l, more preferably 150~250 μ l, still more preferably for
160~200 μ l；
(2) fluorescence of the sample to be tested is measured to detect the presence of the active oxygen or concentration；
Preferably, the sample to be tested is solution or cell, and/or
Preferably, the active oxygen is selected from superoxides, singlet oxygen, hydrogen peroxide, hydroxyl free radical, hypochlorite and peroxide
One or more in nitroso.
7. active oxygen detection method according to claim 6, it is characterised in that when the sample to be tested is solution, step
Suddenly the presence of the detection active oxygen or concentration are carried out by calibration curve method in (2)；
Preferably, the step (2) includes drawing standard curve, draws the standard curve and comprises the following steps：
(a) the fluorescence gold nanoclusters are added into the active oxygen solutions of various concentrations and are reacted,
(b) fluorescence intensity of the active oxygen solutions is detected, is painted according to the concentration of the fluorescence intensity and the active oxygen solutions
Standard curve processed；
It is highly preferred that the concentration of the active oxygen solutions is respectively 0nM, 40nM, 100nM, 200nM, 400nM, 1 μM, 4 μM, 20 μ
M, 40 μM, 60 μM, 80 μM and 250 μM.
8. active oxygen detection method according to claim 6, it is characterised in that when the sample to be tested is cell, step
Suddenly the reaction in (1) is incubation；Preferably, after step (1), the detection method also includes washing away not entering into institute
State the fluorescence gold nanoclusters of cell.
9. a kind of kit, it is characterised in that the kit includes the fluorescence gold nanoclusters described in claim 1 or uses power
Profit requires fluorescence gold nanoclusters prepared by method any one of 2 to 5.
10. method any one of the fluorescence gold nanoclusters or use claim 2 to 5 described in claim 1 prepares glimmering
Kit described in light gold nanoclusters or claim 9 is being prepared for detecting in the product selected from one or more of illness
Application：Cancer, angiocardiopathy, senile dementia and nerve degenerative diseases.
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| CN201610399294.9A CN107478618A (en) | 2016-06-07 | 2016-06-07 | A kind of method of the intracellular ROS detections based on fluorescence gold nanoclusters |
| PCT/CN2016/087357 WO2017210928A1 (en) | 2016-06-07 | 2016-06-27 | Fluorescent gold nanocluster-based method for detecting intracellular ros |
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| CN201610399294.9A CN107478618A (en) | 2016-06-07 | 2016-06-07 | A kind of method of the intracellular ROS detections based on fluorescence gold nanoclusters |
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| CN109091679A (en) * | 2018-09-03 | 2018-12-28 | 国家纳米科学中心 | Gold nano-material, preparation method and application |
| CN109557063A (en) * | 2018-12-30 | 2019-04-02 | 长春中医药大学 | A kind of method by the method for fluorescence copper nano-cluster probe in detecting catecholamine and its concentration, detection monoamine oxidase and its concentration |
| CN114088634A (en) * | 2021-11-18 | 2022-02-25 | 江南大学 | Preparation method of chiral molybdenum selenide nanocluster and application of chiral molybdenum selenide nanocluster in active oxygen detection |
| CN114181171A (en) * | 2021-12-22 | 2022-03-15 | 青岛科技大学 | Fluorescent probe for early diagnosis of Alzheimer's disease |
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| CN111036933A (en) * | 2019-11-26 | 2020-04-21 | 台州学院 | Preparation method and application of supported fluorescent gold nanocluster |
| CN111504961B (en) * | 2020-03-31 | 2023-05-02 | 南昌大学 | Fluorescent phytic acid detection method based on glutathione gold nanoclusters |
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| CN114181171A (en) * | 2021-12-22 | 2022-03-15 | 青岛科技大学 | Fluorescent probe for early diagnosis of Alzheimer's disease |
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