CN104288787B - A kind of long-time cell membrane fluorescence imaging reagent of No clean and preparation method thereof - Google Patents
A kind of long-time cell membrane fluorescence imaging reagent of No clean and preparation method thereof Download PDFInfo
- Publication number
- CN104288787B CN104288787B CN201410487271.4A CN201410487271A CN104288787B CN 104288787 B CN104288787 B CN 104288787B CN 201410487271 A CN201410487271 A CN 201410487271A CN 104288787 B CN104288787 B CN 104288787B
- Authority
- CN
- China
- Prior art keywords
- unit
- glycol
- fluorescence
- chitosan
- cell membrane
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Landscapes
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
A kind of long-time cell membrane fluorescence imaging reagent of No clean that the present invention is provided, the reagent contains the glycol-chitosan macromolecule of hydrophobic units and inefficient unit and fluorescence unit for side chain.The reagent is based on many site grapplings, and its good biocompatibility, to synthesize simple, with low cost, dyeing convenient, without cleaning and being difficult by cell endocytic, can be used as prolonged cell membrane specific trace labelling probe.
Description
Technical field
The invention belongs to biomedicine technical field, more particularly to a kind of long-time based on many site grapplings, No clean
Cell membrane fluorescence imaging reagent, further relates to the preparation method of the reagent.
Background technology
Cell individual is not only wrapped to form cell membrane the internal environment of stabilization in structure, and functionally be take part in
Matter transportation, energy transmission, signal transduction, vesicle transport, cell growth, differentiation, adhesion, transfer, stress, endocytosis, exocytosis, wither
Die, necrosis, many important biomolecule process such as autophagy, therefore the research on cell membrane is increasingly taken seriously.To cell membrane
Carrying out fluorescent staining can be marked and spike to cell membrane, have become the strong hand of research membrane structure and function
Section.
Presently commercially available lipotropism fluorescent dyes such as DiD families (DiO, DiI, DiA, DiR etc.) and FM families (FM4-64,
FM 2-10 etc.) there are many deficiencies.These fluorescence molecules also have stronger fluorescence in aqueous medium, cause excessive fluorescence
Dyestuff is present in cell culture fluid, and background signal interference is greatly, it is necessary to use buffer solution when having in turn resulted in fluorescent microscopic imaging
Or cell culture fluid is cleaned multiple times, satisfied imaging effect could be realized.On the other hand, DiD families are because be
Small molecule, it is easy to dye membrane structure of organelle etc. by endocytosis, therefore lose the specificity of cell membrane imaging.FM contaminates
Although material family improves stability of the dye molecule on cell membrane to a certain extent by way of increasing polar head,
Dyeing still results in endocytosis in more than 10 minutes, it is difficult to mark cell membrane for a long time.In order to solve the problems, such as imaging, mesh for a long time
Preceding also to have many scientific research personnel to develop some new dyes, they are all by the use of one or two hydrophobic tails as grappling
Region, is designed by way of improving hydrophilic head and because be still all small molecule fluorescent dyestuff be easy to by
Endocytosis (when especially dyeing time is long, endocytosis is more very), therefore the mode of this single site grappling has been difficult technological break-through.Cause
This, develops a kind of long-time cell membrane specificity imaging agents that need not be cleaned and is highly desirable to.
Further, since the related biological process of cell membrane needs the tracer study of long time scale mostly, it is therefore desirable to
By long-time cell membrane imaging dyestuff；However, because many biological processes need the state for catching most original in situ (such as
In biological tissue and organ), it is impossible to the operation that fluorescent dye is cleaned commonly used in In vitro cell experiment is carried out, therefore is compeled at present
Being essential will develop the cell membrane imaging dyestuff that a class is capable of No clean.
The content of the invention
Goal of the invention：It is an object of the invention to provide a kind of long-time cell membrane fluorescence imaging reagent of No clean.
Technical scheme：A kind of long-time cell membrane fluorescence imaging reagent of No clean that the present invention is provided, the reagent is
Side chain contains the glycol-chitosan of hydrophobic units and inefficient unit and fluorescence unit.The aqueous solution of the glycol-chitosan in any pH value
In have a good dissolubility, and the features such as with low immunogenicity, biocompatibility, biodegradability, bioactivity.
Hydrophobic units can insert cell membrane by many sites of hydrophobic interaction, and load glycol-chitosan macromolecule and thereon
Nile red molecule anchor on cell membrane；Glycol-chitosan hands over each cholesterol hydrophobic units and Nile red dye unit
Connection gets up, and prevents its dyeing for a long time by cell endocytic；Nile red dye is low due to luminous efficiency in aqueous, and hydrophobic
Cell membrane on send red fluorescence, it is possible to achieve the long-time cell membrane fluorescence imaging of No clean.
Preferably, the hydrophobic units account for the percentage of glycol-chitosan number of repeat unit 5% to 30%；It is described
Nile red dye unit accounts for the percentage of glycol-chitosan number of repeat unit 1% to 5%.
As it is another preferably, the molecular weight of the glycol-chitosan (glycol chitosan) more than 10000,
It is preferred that 10000-100000.
As another kind preferably, the hydrophobic units include cholesterol, phospholipid molecule, alkane and vitamin E；It is described hydrophobic
Unit is linked on the amino of glycol-chitosan by polyethylene glycol polymer；The wherein polyethylene glycol polymer molecular weight
More than 500.
As another kind preferably, the inefficient unit and fluorescence unit is the Nile red dye of carboxylated or the tetraphenyl of carboxylated
Ethene, it is directly linked on glycol-chitosan by with amino reaction, or is linked to glycol-chitosan by short chain PEG
Amino on.Luminous efficiency is low in aqueous for the inefficient unit and fluorescence unit, and luminous intensity is big in hydrophobic environment
Fluorescence molecule, to realize that No clean is imaged.
The structural formula of the Nile red dye of the carboxylated is：
The structural formula of the tetraphenylethylene of the carboxylated is：
Present invention also offers a kind of preparation method of the long-time cell membrane fluorescence imaging reagent of above-mentioned No clean, including
Following steps：
(1) take hydrophobic units molecule and glycol-chitosan is dissolved in PBS respectively, mix, room temperature reaction；Product
Dialyse, freeze；
(2) it is sub- in 1- ethyls-(3- dimethylaminopropyls) carbodiimide hydrochloride (EDC) and N- hydroxies succinyl
In the presence of amine (sulfo-NHS), product is obtained final product with inefficient unit and fluorescence unit molecule room temperature reaction, purifying.
Beneficial effect：The present invention provide cell membrane fluorescence imaging reagent be based on many site grapplings, its good biocompatibility,
Synthesis is simple, with low cost, dyeing is convenient, without cleaning and being difficult by cell endocytic, can be used as prolonged cell membrane special
Different in nature trace labelling probe.
Specifically, it is of the invention relative to prior art, with advantage following prominent：
(1) present invention employs many site anchoring techniques, as long as there is a cholesterol molecule insertion thin on macromolecule
After after birth, the cholesterol of other anchor units on same macromolecular scaffold will be promoted to insert film successively and (to be existed and inserted membrane process
Cooperative effect), it is short the time required to fluorescent staining.
(2) hydrophobic units of reagent of the present invention and inefficient unit and fluorescence unit are all linked at ethylene glycol in covalently bound mode
On the skeleton of shitosan macromolecule, resulting imaging agents can carry out many site insertions with cell membrane, form high score attached bag
By structure, it is set to be difficult by cell endocytic, therefore increased the retention time on cell membrane.
(3) the inefficient unit and fluorescence unit of reagent of the present invention luminous efficiency in water is low, and inserts the meeting after cell membrane and dredging
Send fluorescence in the phospholipid layer of water, even if therefore be not inserted into cell membrane free dye do not wash there will not be background do
Disturb, it is achieved thereby that No clean is imaged；Simultaneously as dyestuff is difficult endocytosis, so free dyestuff is constantly fed to film again
On, it is achieved that prolonged fluorescence imaging.
(4) constituent of the reagent includes that the selection of glycol-chitosan and PEG all possesses biocompatibility, Er Qiemao
The cholesterol for determining side chain is even more zooblast institute proper constituent, therefore the reagent toxicity is low, and biological cell membrane marker will not be produced
Raw influence.
(5) glycol-chitosan and PEG of the reagent make the imaging agents have water solubility well, can easily enter
Row In vitro cell experiment and living imaging are tested.
(6) positive charge that remaining amino is carried on the reagent glycol-chitosan, can send out with negatively charged cell membrane
Raw electrostatic interaction, increases the affinity with cell membrane, thus further promotes retention time of the dyestuff on cell membrane.
(7) many site anchoring schemes proposed by the invention can be as an application platform, by changing different anchor units, such as
Using the stronger unit and fluorescence unit of fluorescence under the hydrophobic environments such as tetraphenylethylene (TPE), such that it is able to develop various functions and each
The probe of color is planted, more complicated and special membrane structure and functional study is applied to.
Brief description of the drawings
Fig. 1 be based on many site grapplings, No clean long-time cell membrane fluorescence imaging reagent pie graph (fluorescence molecule with
As a example by Nile red molecule).
Fig. 2 is the Confocal microscopy imaging results to A549 cells using fluorescence imaging reagent.Fluorescence imaging reagent is
Chitosan-30%cholesterol-2%Nile red, its preparation process is shown in embodiment 1.
Specific embodiment
Embodiment 1
Many site grapplings, the long-time cell membrane fluorescence imaging reagent chitosan-30%cholesterol- of No clean
The design of 2%Nile red, synthesis and cell membrane fluorescent staining method are as follows：
The design of reagent：The reagent is constituted sees Fig. 1, and the reagent is with glycol-chitosan macromolecule (glycol
Chitosan it is) skeleton, side chain contains 30% polyethylene glycol 2000-cholesterol (PEG2000-cholesterol) hydrophobic list
Unit and 2% Nile red dye unit and fluorescence unit.Described glycol-chitosan macromolecule, with good biocompatibility and water
Dissolubility, molecular weight is more than 10000.Described cholesterol hydrophobic side chain, by the form of NHS-PEG2000-cholesterol
It is linked on the amino of glycol-chitosan, increases water-soluble.Described Nile red dye for carboxylated Nile red, by with
Amino reaction is directly linked on glycol-chitosan.After the reagent is dissolved in water, free extended position is kept, will not self assembly
Into nano particle, it is difficult by cell endocytic, realizes being imaged for a long time.Cholesterol hydrophobic units can be by hydrophobic interaction multidigit
The insertion cell membrane of point, and macromolecule and Nile red molecule are anchored on cell membrane；Macromolecule is the hydrophobic list of each cholesterol
Unit and the crosslinking of Nile red dye unit are got up, and prevent its dyeing for a long time by cell endocytic；Nile red dye is due in the aqueous solution
Middle luminous efficiency is low, and the fluorescence of red is sent on hydrophobic cell membrane, realize the long-time cell membrane fluorescence of No clean into
Picture.
The synthesis of reagent：It is sub- with glycol-chitosan macromolecule (glycol chitosan), N- hydroxysuccinimidyls acyl first
Amine-polyethylene glycol 2000-cholesterol (NHS-PEG2000-cholesterol) and Nile red dye are raw material, synthesize grappling list
The percentage that unit accounts for glycol-chitosan unit number is 30%, and fluorescence molecule unit accounts for the percentage of glycol-chitosan unit number
Be 2% imaging agents, i.e. chitosan-30%cholesterol-2%Nile red dyestuffs, specially：5mg is weighed respectively
N-hydroxy-succinamide-polyethylene glycol 2000-cholesterol and the 1.34mg glycol-chitosan macromolecule (degree of polymerization：It is more than
400；80000) molecular weight about and is dissolved in PBS (pH 7.4,50mM) respectively, then after both are mixed, magneton teeter chamber
Temperature reaction 4h；Reaction terminates to be made centre in the bag filter of 10k, dialysis is purified and freezed for 3 days after molecular cut off
Product " glycol-chitosan-cholesterol " compound；Then, 1mg " glycol-chitosan-cholesterol " and 2nmol carboxyls are weighed
(structural formula is shown in Fig. 2 to the Nile red dye of change；Synthetic method sees reference document：Benzophenoxazine-based
Fluorescent dyes for labeling biomolecules.Tetrahedron 2006,62,11021-11037) in
In DMSO, the NHS (being 3 times of excess) of the EDC and 6nmol of 6nmol is added in room temperature reaction overnight；Reaction is used after terminating
HPLC methods are purified, most after the final obtained imaging agents chitosan-30% of freeze-drying in freeze dryer
Cholesterol-2%Nile red dyestuffs are in freezen protective in -20 DEG C.
Cell membrane dyes imaging experiment：The imaging agents chitosan-30%cholesterol-2%Nile red dyes
Material is dissolved in the storing liquid that cell culture is made 4mg/mL of PBS, then with DMEM complete mediums, (DMEM is not exclusively trained by storing liquid
Support+1% Pen .- Strep of hyclone of base+10% dual anti-) it is diluted to the working solution of final concentration of 100 μ g/mL.Cell exists
After being cultivated 12 hours in copolymerization Jiao's culture plate, culture medium is first sucked, PBS is washed twice, adds 100 μ g/mL working solutions to put back to cell
Incubator dyes 5min.Dyeing need not be cleaned after terminating, directly in Nile red dye fluorescence under laser scanning co-focusing microscope
Passage is shown in Fig. 2 in 63 times of oily Microscopic observations, its imaging results.
Embodiment 2
The implementation of the present embodiment is consistent with the method in embodiment 1, simply changes Nile red fluorescence molecule into four benzene
Base ethene (tetraphenylethylene, TPE) fluorescence molecule, the glycol-chitosan molecular weight polymeric of selection is about
100000.The fluorescence molecule is one kind of aggregation-induced emission (AIE), and it can be limited (restriction by Internal Rotations of Molecules
Of intra-molecular rotation, RIR) mode light.The fluorescence molecule in aqueous when the free degree it is big, energy
Amount easily dispersion, so luminous efficiency is low；And after hydrophobic cell membranes are inserted, the sky of the deflation due to being bound in hydrophobic phospholipid
Between in, molecule limited swivel, power dissipation is obstructed, produce high-fluorescence quantum yield green fluorescence.So the fluorescence molecule
Luminescent condition is similar to Nile red, can be used for being made the long-time cell membrane fluorescence imaging reagent of many site grapplings, No clean.
In preparation scheme, by tetraphenylethylene molecule TPE-COOH (the synthetic method bibliography of carboxylated：Cell Membrane
Tracker Based on Restriction of Intramolecular Rotation.ACS
Appl.Mater.Interfaces, 2014,6,8971-8975) second two is linked to by amido link under the catalysis of EDC and NHS
On the macromolecular scaffold of alcohol shitosan.
Embodiment 3
The implementation of the present embodiment is similar with the method in embodiment 1, the glycol-chitosan macromolecule simply selected
Molecular weight about 10000, the percentage that synthesis anchor unit accounts for glycol-chitosan number of repeat unit is changed to 5%, fluorescence molecule list
The percentage that unit accounts for glycol-chitosan number of repeat unit is changed to 1%, and obtained imaging agents composition is chitosan-5%
Cholesterol-1%Nile red.
Embodiment 4
The implementation of the present embodiment is similar with the method in embodiment 1, and simply synthesis anchor unit accounts for ethylene glycol shell and gathers
The percentage of sugared number of repeat unit is changed to 20%, and the percentage that fluorescence molecule unit accounts for glycol-chitosan number of repeat unit is changed to
5%, obtained imaging agents composition is chitosan-20%cholesterol-5%Nile red.
Embodiment 5
The implementation of the present embodiment is consistent with the method in embodiment 1, simply NHS-PEG2000-cholesterol points
Cholesterol hydrophobic units in son make phospholipid molecule (1,2- myristoyl phosphatidyl-ethanolamines, DMPE), alkane (ten into successively
Eight carbon atoms, C18) or the hydrophobic side chain such as vitamin E cell membrane fluorescence imaging reagent is obtained.
Claims (6)
1. the long-time cell membrane fluorescence imaging reagent of a kind of No clean, it is characterised in that：The reagent contains film anchor for side chain
Order unit and the glycol-chitosan of inefficient unit and fluorescence unit, wherein, film anchor unit is by hydrophobic units and polyethylene glycol high score
Son composition, hydrophobic units are linked on the amino of glycol-chitosan by polyethylene glycol polymer；Inefficient unit and fluorescence unit is
Luminous efficiency is low in aqueous, and the big fluorescence molecule of luminous intensity in hydrophobic environment.
2. the long-time cell membrane fluorescence imaging reagent of a kind of No clean according to claim 1, it is characterised in that：It is described
Film anchor unit accounts for the percentage of glycol-chitosan number of repeat unit 5% to 30%；The inefficient unit and fluorescence unit accounts for second
The percentage of glycol shitosan number of repeat unit is 1% to 5%.
3. the long-time cell membrane fluorescence imaging reagent of a kind of No clean according to claim 1, it is characterised in that：It is described
The molecular weight of glycol-chitosan is 10000-100000.
4. the long-time cell membrane fluorescence imaging reagent of a kind of No clean according to claim 1, it is characterised in that：It is described
Film anchor unit includes cholesterol, phospholipid molecule, alkane and vitamin E；The hydrophobic units pass through polyethylene glycol polymer chain
It is connected on the amino of glycol-chitosan, the polyethylene glycol polymer molecular weight is more than 500.
5. the long-time cell membrane fluorescence imaging reagent of a kind of No clean according to claim 1, it is characterised in that：It is described
Inefficient unit and fluorescence unit is the Nile red dye of carboxylated or the tetraphenylethylene of carboxylated, and it reacts direct chain by with amino
It is connected on glycol-chitosan, or is linked on the amino of glycol-chitosan by short chain PEG.
6. the preparation method of the long-time cell membrane fluorescence imaging reagent of a kind of No clean, it is characterised in that comprise the following steps：
(1) take film anchor unit and glycol-chitosan is dissolved in PBS respectively, mix, room temperature reaction；Product dialysis,
It is lyophilized；
(2) in the presence of 1- ethyls-(3- dimethylaminopropyls) carbodiimide salt and N- hydroxy thiosuccinimides, product
With inefficient unit and fluorescence unit molecule room temperature reaction, purifying obtains final product；Wherein, the inefficient unit and fluorescence unit is luminous in aqueous
Efficiency is low, and the big fluorescence molecule of luminous intensity in hydrophobic environment.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410487271.4A CN104288787B (en) | 2014-09-22 | 2014-09-22 | A kind of long-time cell membrane fluorescence imaging reagent of No clean and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410487271.4A CN104288787B (en) | 2014-09-22 | 2014-09-22 | A kind of long-time cell membrane fluorescence imaging reagent of No clean and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104288787A CN104288787A (en) | 2015-01-21 |
| CN104288787B true CN104288787B (en) | 2017-06-09 |
Family
ID=52308892
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410487271.4A Expired - Fee Related CN104288787B (en) | 2014-09-22 | 2014-09-22 | A kind of long-time cell membrane fluorescence imaging reagent of No clean and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104288787B (en) |
Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104888219B (en) * | 2015-06-10 | 2017-11-03 | 东南大学 | A kind of tumour phototherapy reagent coated based on cell membrane and its preparation method and application |
| CN105969334B (en) * | 2016-05-06 | 2018-07-20 | 东南大学 | Fluorescent dyeing reagent, Preparation method and use for detection bacterium and fungi life or death state |
| CN106957401B (en) * | 2017-04-01 | 2020-03-17 | 苏州大学 | Cholesterol group anchored poly (ethylene glycol) methacrylate polymer and synthesis method and application method thereof |
| CN107941575B (en) * | 2017-11-06 | 2020-03-31 | 东南大学 | Washing-free cell membrane red fluorescence imaging reagent and preparation method and application thereof |
| CN109852379A (en) * | 2019-03-13 | 2019-06-07 | 中国科学院苏州纳米技术与纳米仿生研究所 | The area near-infrared II fluorescence quantum cell membrane tagging system, labeling method and application |
| CN110836879B (en) * | 2019-11-06 | 2021-12-07 | 东南大学 | Cell membrane long-time multicolor fluorescence imaging reagent and preparation method and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102973948A (en) * | 2012-12-03 | 2013-03-20 | 上海交通大学 | Method for preparing drug carrier based on magnetic carbon quantum dot/chitosan composite microsphere |
| CN103087328A (en) * | 2013-02-01 | 2013-05-08 | 北京理工大学 | Preparation method of chitosan 6-OH immobilized cyclodextrin derivative and application thereof in H2O2 detection fluorescent biosensor |
-
2014
- 2014-09-22 CN CN201410487271.4A patent/CN104288787B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102973948A (en) * | 2012-12-03 | 2013-03-20 | 上海交通大学 | Method for preparing drug carrier based on magnetic carbon quantum dot/chitosan composite microsphere |
| CN103087328A (en) * | 2013-02-01 | 2013-05-08 | 北京理工大学 | Preparation method of chitosan 6-OH immobilized cyclodextrin derivative and application thereof in H2O2 detection fluorescent biosensor |
Non-Patent Citations (1)
| Title |
|---|
| "A new atherosclerotic lesion probe based on hydrophobically modified chitosan nanoparticles functionalized by the atherosclerotic plaque targeted peptides";Kyeongsoon Park等;《Journal of Controlled Release》;20081231;第128卷;摘要，第2部分 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104288787A (en) | 2015-01-21 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104288787B (en) | A kind of long-time cell membrane fluorescence imaging reagent of No clean and preparation method thereof | |
| Feng et al. | Conjugated polymer nanoparticles: preparation, properties, functionalization and biological applications | |
| Ma et al. | Hyaluronic acid-conjugated mesoporous silica nanoparticles: excellent colloidal dispersity in physiological fluids and targeting efficacy | |
| CN105169474B (en) | Polypeptide material capable of carrying out self-assembly to form hydrogel under neutral pH condition and applications thereof | |
| US20080051646A1 (en) | Probe for cellular oxygen | |
| CN110237035B (en) | Active targeting amphiphilic polypeptide nano-drug carrier and preparation and application thereof | |
| Jiang et al. | The effects of an RGD-PAMAM dendrimer conjugate in 3D spheroid culture on cell proliferation, expression and aggregation | |
| CN104093768B (en) | It is imported with block copolymer and the use thereof of phenylboric acid base | |
| CN106085416A (en) | A kind of aggregation-induced emission nanoparticle and its preparation method and application | |
| Niu et al. | A new AIE multi-block polyurethane copolymer material for subcellular microfilament imaging in living cells | |
| CN104231114B (en) | A kind of cell membrane fluorescence imaging reagent based on the grappling of many sites and preparation method thereof | |
| TW201524533A (en) | Polymeric micelle pharmaceutical composition | |
| CN100562341C (en) | The application of cell nucleus targeting chitosan-fatty acid graft as medicine carrier micelle | |
| Wang et al. | Ultra long-term cellular tracing by a fluorescent AIE bioconjugate with good water solubility over a wide pH range | |
| Huang et al. | Lipoic acid based core cross-linked micelles for multivalent platforms: design, synthesis and application in bio-imaging and drug delivery | |
| CN111138682A (en) | Fluorescence-labeled biological material and preparation method thereof | |
| US20230003727A1 (en) | Luminescent zwitterionic polymeric nanoparticles | |
| CN102961756A (en) | Synthesis method of polypeptide-nanogold particle drug carrier | |
| KR101531046B1 (en) | Surface-modified nanofiber, a process for the preparation thereof, and a use thereof | |
| Teng et al. | In vitro characterization of pH-sensitive azithromycin-loaded methoxy poly (ethylene glycol)-block-poly (aspartic acid-graft-imidazole) micelles | |
| CN105168130A (en) | Tumor-targeted polymer micelle and preparation method thereof | |
| CN104644560A (en) | Hyaluronic acid modified hectorite amide nanoparticle and preparation and application of hyaluronic acid modified hectorite amide nanoparticle | |
| CN110836879B (en) | Cell membrane long-time multicolor fluorescence imaging reagent and preparation method and application thereof | |
| CN104316500A (en) | Long-time cell membrane imaging agent and preparation method thereof | |
| Chen et al. | Dual fluorescence nano-conjugates based on gold nanoclusters for tumor-targeting imaging |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20170609 Termination date: 20190922 |