CN104892815A - Luminescent nano micro-spheres with positive charge on surface and possessing aggregation induced fluorescence enhancement property and biological application thereof - Google Patents
Luminescent nano micro-spheres with positive charge on surface and possessing aggregation induced fluorescence enhancement property and biological application thereof Download PDFInfo
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Abstract
The invention relates to luminescent nano micro-spheres with positive charge on surface and possessing aggregation induced fluorescence enhancement property and a biological application thereof, which belongs to the technical field of high-molecular material. According to the invention, the nano micro-spheres with positive charge on surface is synthesized, the fluorescence molecules with positive charge and AIE effect are modified to the surface of the nano micro-spheres through electrostatic interactions. Molecule internal rotation of fluorescence molecule is restricted by Coulombian force effect, the absorbed energy can be basically released through fluorescence radiation, accordingly fluorescence from the fluorescence molecule modified to the nano micro-spheres can be enhanced by a hundred times, and excellent AIE property can be generated. The prepared the luminescent nano micro-spheres have the advantages of stable fluorescence property, good biological compatibility and low toxicity, and the positive charge on the surface is easily introduced to cell and biology detection. The luminescent nano micro-spheres with positive charge on surface has wide application prospect on the biology fields such as cell imaging.
Description
Technical field
The invention belongs to technical field of polymer materials, be specifically related to a kind of surface band positive charge and there is the fluorescent nanometer microsphere of aggregation inducing Fluorescence Increasing character and the application in cell imaging and biological detection etc. thereof.
Background technology
Bio-imaging is set multiple technologies, the multiple subject that blends, is widely used, develops emerging field rapidly.In the last few years along with the development of biological chemistry and life science, people had turned to microcosmic from macroscopic view gradually to the research of organism.Therefore the bioluminescence imaging in one of its branch field becomes the focus of people's research and concern.Fluorescence technique has quick, sensitive, real-time, "dead", reproducible, and multiple optical physics parameter (as emission wavelength, excitation wavelength, fluorescence intensity, fluorescence lifetime) can be used for the advantages such as detection.Fluorescent probe is the core technology realizing bioluminescence imaging.Traditional fluorescent probe based on organic fluorescent dye, but because organic fluorescent dye photostabilization is poor, poorly water-soluble, poor biocompatibility and assemble the shortcomings such as fluorescence induction cancellation, and be difficult to apply in practice.Increasingly extensive along with bioluminescence imaging applications, develops novel fluorescent probe and becomes very urgent with the shortcoming overcoming conventional probe.Therefore, fluorescent nano particles probe is suggested as the effective means addressed this problem.
Current fluorescent nano particles probe mainly comprises: quantum dot (quantum dot, QD), upper conversion rare-earth nanometer particles (Upconversion Rare Earth Nanoparticles, UCRE-NPs) and high molecular fluorescent Nano microsphere.Inorganic-quantum-dot is widely used in the fields such as biomarker because fluorescence efficiency is high by people, but research finds when inorganic-quantum-dot enters in organism for bio-imaging, oxidizing one-tenth heavy metal ion in the easy body in surface, therefore bio-toxicity is unsuitable for bio-imaging greatly.High molecular fluorescent Nano microsphere starts to be with polystyrene, polyacrylamide, polymethacrylate for particulate body, the fluorescent nanometer microsphere of surface bond or absorption fluorescent substance.Because single nanoparticle can the multiple fluorescence molecule of bonding, so fluorescence intensity strengthens to some extent.Compared with micromolecular organic dye, high molecular fluorescent Nano microsphere usually in aqueous phase comparatively strong, the luminous efficiency of favorable dispersity, fluorescent emission intensity higher while, the phenomenon occurrence probability of photobleaching is very low; And particle stability and biocompatibility are all very high under multiple coenocorrelation.So a lot of high molecular fluorescent nanoparticle is without the need to being just directly used in the biologic applications fields such as bio-imaging fluorescent probe, biological sensor through loaded down with trivial details modification and improvement.But the most of fluorescence molecule at present for high molecular fluorescent Nano microsphere has aggregation inducing Quenching.When preparing high-level efficiency high molecular fluorescent Nano microsphere, fluorescence dye add-on is few, fluorescent signal can not improve significantly, but also can decay due to aggregation inducing quenching effect fluorescent signal when add-on is large or not have fluorescent emission, makes it apply and is restricted.
In recent years, the fluorescence molecule with aggregation-induced emission (AIE) receives the concern of people.It is different from traditional fluorescence molecule, and AIE molecule fluorescence in state of aggregation or poor solvent can significantly strengthen, and makes fluorescence decay when being in free state because internal rotation aggravation consumes the energy absorbed.But AIE molecule mostly is organic dye small molecules, poor biocompatibility, cycling time, therefore the short cell that can not effectively enter can not be used for bio-imaging in blood.Therefore design effective carrier and can realize the AIE effect of fluorescence molecule and good biocompatibility, bio-toxicity is low, and size is suitable, and the good and internal milieu of Cell permeable does not have cancellation effect to its fluorescence, is vital in bio-imaging field.
Summary of the invention
The object of this invention is to provide a kind of fluorescent nanometer microsphere with aggregation inducing Fluorescence Increasing character that can be used for the surface band positive charge of cell imaging and biological detection.
First the present invention synthesizes a kind of Nano microsphere emulsion of surface band positive charge, and the fluorescence molecule with AIE effect with negative charge is modified Nano microsphere surface by electrostatic force.Fluorescence molecule is restricted due to the effect internal rotation being subject to Coulomb's force, and the energy of absorption discharges essentially by fluorescent radiation, and therefore fluorescence molecule is modified its fluorescence intensity on Nano microsphere and strengthened hundred times, shows excellent AIE character.Fluorescent nanometer microsphere photoluminescent property obtained by us is stablized, and good biocompatibility, toxicity is low, and surface band positive charge is easy to enter cell interior, carries out cell fluorescence imaging and biological detection.Therefore, the fluorescent nanometer microsphere of surface band positive charge that we obtain has broad application prospects at biological fields such as cell imagings.
The fluorescent nanometer microsphere with aggregation inducing Fluorescence Increasing character of surface band positive charge of the present invention, it is prepared by following steps:
(1) measure 5 ~ 10mL polymerization single polymerization monomer 1 to be scattered in the deionized water of 150 ~ 200mL, then add the polymerization single polymerization monomer 2 of 0.04g ~ 0.5g; Under room temperature, nitrogen protection, mechanical stirring (300 ~ 600rpm), the oxygen in removing reaction system; Add the aqueous solution of 10mL containing 0.3 ~ 0.5mmol initiator after being warming up to 70 ~ 90 DEG C in reaction system, nitrogen protection, the lower polyreaction of stirring 5 ~ 20 hours, obtain the Nano microsphere solution of surface band positive charge; Impurity such as high speed centrifugation (15000 ~ 20000rpm) washing removing unreacted monomer, oligopolymer, initiator etc., Nano microsphere again ultrasonic disperse in 100mL deionized water;
(2) the Nano microsphere solution that 10 ~ 20mL step (1) obtains is measured, add 10 ~ 20mL deionized water, and then add the AIE type fluorescence molecule of 3 ~ 5mL, 80 ~ 150 μ g/mL, 50 ~ 80 DEG C, under nitrogen protection condition, magnetic agitation reaction 4 ~ 10h; High speed centrifugation (15000 ~ 20000rpm) removes the AIE type fluorescence molecule of non-compound, obtains the fluorescent nanometer microsphere of the surface band positive charge of AIE molecule compound; The fluorescent nanometer microsphere obtained is scattered in deionized water, thus obtains the fluorescent nanometer microsphere with aggregation inducing Fluorescence Increasing character of surface band positive charge of the present invention.
In aforesaid method, polymerization single polymerization monomer 1 is vinylbenzene (St), fluorostyrene (F-St), methyl methacrylate (MMA), β-dimethyl-aminoethylmethacrylate, ethyl propenoate, vinylchlorid, tert-butyl acrylate, Vinylstyrene or alpha-methyl styrene or glycidyl methacrylate (GMA);
In aforesaid method, polymerization single polymerization monomer 2 is N, N, N-trimethyl-ethylene base benzene first ammonium chloride (VBTAC), 2-(diisopropylaminoethyl) β-dimethyl-aminoethylmethacrylate (DPA) or 2-aminoethyl methacrylate hydrochloride (AMA);
In aforesaid method, initiator is azo-bis-isobutyrate hydrochloride (V
50), azo two isobutyl imidazoline hydrochloride (AIBI), azo isobutyl cyano group methane amide (V
30);
In aforesaid method, the fluorescence molecule of AIE type is 9,10-bis-(styryl) anthracene sulfonic acid salt (DSA), 1,2-bis-[4-(3-sulfonic acid propoxy-) phenyl]-1,2-diphenylethlene disodium salt (BSTPE) or 1,1,2,2-tetra-[4-(3-sulfonic acid propoxy-) phenyl] ethene tetra-na salt (TSTPE).
Tool of the present invention has the following advantages:
1, the particle diameter of this fluorescent nanometer microsphere is at Nano grade and size controlled (50 ~ 140nm);
2, this fluorescent nanometer microsphere good biocompatibility, energy stable existence more than 3 months (Fig. 5) in phosphate buffer solution and biological macromolecule solns;
3, this fluorescence molecule fluorescence in the scope of pH=3 ~ 7 is basicly stable, and in the scope of pH=7 ~ 10, fluorescence linearly reduces, but reduction degree does not affect fluorescence imaging;
4, this fluorescent nanometer microsphere surface band positive charge, is easy to enter cell, carries out cell fluorescence imaging and biological detection.
5, this fluorescent nanometer microsphere is biomacromolecule as fluorescent signal when BSA exists strengthens, and biomacromolecule can play the effect of amplifying fluorescent signal.And BSA concentration and fluorescence intensity linearly change, therefore this material can be used for detecting BSA concentration (Fig. 7).
6, the cytotoxicity of this fluorescent nanometer microsphere low (Fig. 8), is beneficial to it and carries out biologic applications.
Accompanying drawing explanation
Fig. 1: the Nano microsphere electron scanning micrograph (SEM) prepared for embodiment 1;
Fig. 2: the Nano microsphere prepared for embodiment 1 and fluorescent nanometer microsphere dynamic light scattering grain-size graph;
Fig. 3: the Nano microsphere prepared for embodiment 1 and fluorescent nanometer microsphere dynamic light scattering Zeta scheme;
Fig. 4: the fluorescent nanometer microsphere prepared for embodiment 1 and fluorescence molecule aqueous solution fluorogram (fluorescence molecule concentration is identical, λ ex=420nm);
Fig. 5: the dynamic light scattering grain-size graph of the fluorescent nanometer microsphere aqueous solution prepared for embodiment 1, fluorescent nanometer microsphere foetal calf serum (FBS) phosphate buffer solution (pH=7.4), fluorescent nanometer microsphere foetal calf serum (FBS) phosphate buffer solution (pH=7.4) after 3 months;
Fig. 6: the fluorescent nanometer microsphere that (1) prepares for embodiment 1 under different pH phosphate buffer solution, the fluorogram of fluorescent nanometer microsphere; (2) be fluorescent nanometer microsphere fluorescence intensity and the pH graph of a relation of embodiment 1 preparation;
Fig. 7: Nano microsphere fluorogram under different B SA concentration that (1) prepares for embodiment 1; (2) be BSA concentration and fluorescence intensity curves figure;
Fig. 8: (1) is for embodiment 1 fluorescent nanometer microsphere is to the cytotoxicity test pattern of people's gastric epithelial cell (GES-1); (2) for embodiment 1 fluorescent nanometer microsphere is to the cytotoxicity test pattern of gastric carcinoma cells (SGC-7901);
Fig. 9: the Nano microsphere that (a) is prepared for embodiment 1 is at normal cell people gastric epithelial cell (GES-1) fluorescence co-focusing photograph via bright field; B Nano microsphere that () prepares for embodiment 1 superposes photo at normal cell people gastric epithelial cell (GES-1) fluorescence co-focusing light field and details in a play not acted out on stage, but told through dialogues; C Nano microsphere that () prepares for embodiment 1 is at normal cell people gastric epithelial cell (GES-1) fluorescence co-focusing details in a play not acted out on stage, but told through dialogues photo; D Nano microsphere that () prepares for embodiment 1 is at normal cell gastric carcinoma cells (SGC-7901) fluorescence co-focusing photograph via bright field; E Nano microsphere that () prepares for embodiment 1 superposes photo at gastric carcinoma cells (SGC-7901) fluorescence co-focusing light field and details in a play not acted out on stage, but told through dialogues; F Nano microsphere that () prepares for embodiment 1 is at gastric carcinoma cells (SGC-7901) fluorescence co-focusing details in a play not acted out on stage, but told through dialogues photo;
Figure 10: the Nano microsphere electron scanning micrograph (SEM) prepared for embodiment 2;
Figure 11: the Nano microsphere electron scanning micrograph (SEM) prepared for embodiment 3;
Figure 12: the Nano microsphere electron scanning micrograph (SEM) prepared for embodiment 4.
Embodiment
Embodiment 1:
(1) 5mL vinylbenzene (analytical pure is got; through underpressure distillation except stopper) and 0.06g N; N; N-trimethyl-ethylene base benzene first ammonium chloride (VBTAC) joins in the three-necked bottle of the 500mL containing 185mL deionized water; mechanical stirring (400rpm) 30 minutes under room temperature, nitrogen protection; oxygen in removing reaction system, is then warmed up to 70 DEG C, adds 10mL, containing 0.37mmol azo-bis-isobutyrate hydrochloride (V
50) aqueous solution initiated polymerization of initiator, carry out 10h under being aggregated in the stirring velocity of nitrogen protection, 400rpm.Be polymerized the Nano microsphere that obtains under the rotating speed of 18500rpm centrifugal for 3 times, and with deionized water wash 3 times, remove unreacted monomer, oligopolymer, initiator etc., be re-dispersed in 100mL deionized water, just obtained mass concentration is the Nano microsphere solution of 2.92% surface band positive charge.
(2) get 10mL Nano microsphere solution to add 10mL deionized water and add 3mL, 100 μ g/mL DSA again in three-necked bottle, put into stirrer, 50 DEG C of reaction 4h are heated under magnetic stirs, reacted rear fluorescent nanometer microsphere under the rotating speed of 18500rpm centrifugal for 3 times, and with deionized water wash 3 times, remove the fluorescence molecule of non-compound, and be re-dispersed in 23mL deionized water, obtain the fluorescent nanometer microsphere solution with aggregation inducing Fluorescence Increasing character that mass concentration is the surface band positive charge of 1.27%.
Can find out that Nano microsphere is of a size of 100nm from Nano microsphere electron scanning micrograph (SEM) (Fig. 1), size uniformity, monodispersity is good.Record Nano microsphere by dynamic light scattering (DLS) and be of a size of 120.7nm in aqueous, single dispersing indices P DI=0.002 (Fig. 2), owing to there is hydration particle diameter, so the size recorded is larger than the numerical value of SEM.Nano microsphere surface is recorded in positive polarity (42.4mv) (Fig. 3) by dynamic light scattering (DLS).Electronegative AIE type fluorescence molecule DSA is compound to fluorescent nanometer microsphere surface by the effect of Coulomb's force, the hydration particle diameter of the fluorescent nanometer microsphere obtained is 131.7nm (Fig. 2), and there is good monodispersity, single dispersing indices P DI=0.013, surface charge is also reduced to 40.6mv (Fig. 3).Although fluorescence molecule DSA itself has good biocompatibility, and overcome traditional gathering fluorescence induction quenching phenomenon, but water-soluble owing to having, and most of energy of absorption is all discharged by internal rotation, fluorescence efficiency is very low, is not therefore suitable for bio-imaging.Fluorescence molecule DSA be compound to we synthesis Nano microsphere on after, because internal rotation is restricted, the energy that major part absorbs is discharged by the mode of radiation, can find out the enhancing significantly (Fig. 4) that fluorescence efficiency obtains from fluorogram.Fluorescent nanometer microsphere synthesized by us has good biocompatibility and biologically stable.We have prepared 10% foetal calf serum (FBS) phosphate buffer solution (pH=7.4) containing 500 μ g/mL fluorescent nanometer microspheres, low-temp storage, after 3 months, records size by dynamic light scattering and considerable change (Fig. 5) does not occur.And we test the fluorescent stability of fluorescent nanometer microsphere under different coenocorrelation, when fluorescent nanometer microsphere is in different pH phosphate buffer solutions, fluorescence intensity is significantly change (Fig. 6) not.When in biomacromolecule BSA solution, because these biomacromolecules are electronegative, the surface being adsorbed onto fluorescent nanometer microsphere fetters DSA internal rotation further, therefore make its Fluorescence Increasing and linearly change (Fig. 7), therefore more intracellular biomolecules can amplify the fluorescent signal of the fluorescent nanometer microsphere of our synthesis.We conducted the cytotoxicity test of fluorescent nanometer microsphere, can find out that the survival rate of cultivating 72h cell at Nano microsphere concentration 100mg/L still reaches more than 85%, cytotoxicity is low, may be used for bio-imaging (Fig. 8).We have carried out cell imaging experiment to it, see that fluorescent nanometer microsphere that we synthesize can enter the tenuigenin of normal cell and cancer cells well, carry out good imaging (Fig. 9) to cell by the burnt cell imaging of copolymerization.
Embodiment 2:
(1) 5mL vinylbenzene (analytical pure is got; through underpressure distillation except stopper) and 0.5g N; N; N-trimethyl-ethylene base benzene first ammonium chloride (VBTAC) joins in the three-necked bottle of the 500mL containing 185mL deionized water; mechanical stirring (400rpm) 30 minutes under room temperature, nitrogen protection; oxygen in removing reaction system, is then warmed up to 70 DEG C, adds 10mL, containing 0.37mmol azo-bis-isobutyrate hydrochloride (V
50) aqueous solution initiated polymerization of initiator, carry out 10h under being aggregated in the stirring velocity of nitrogen protection, 400rpm.Be polymerized the Nano microsphere that obtains under the rotating speed of 18500rpm centrifugal for 3 times, and with deionized water wash 3 times, remove unreacted, oligopolymer, initiator etc., be re-dispersed in 100mL deionized water, just obtained mass concentration is the Nano microsphere solution of 2.92% surface band positive charge.
(2) get 10mL Nano microsphere solution to add 10mL deionized water and add 3mL, 100 μ g/mL DSA again in three-necked bottle, put into stirrer, 50 DEG C of reaction 4h are heated under magnetic stirs, reacted rear fluorescent nanometer microsphere under the rotating speed of 18500rpm centrifugal for 3 times, and with deionized water wash 3 times, remove the fluorescence molecule of non-compound, and be re-dispersed in 23mL deionized water, obtain the fluorescent nanometer microsphere solution with aggregation inducing Fluorescence Increasing character that mass concentration is the surface band positive charge of 1.27%.
We find, its character is similar with the properties of samples of preparation in embodiment 1, but size diminishes, and is about 50nm (see Figure 10).This is because polymerization single polymerization monomer N, N, N-trimethyl-ethylene base benzene first ammonium chloride (VBTAC) is amphiphilic monomer, serves as the effect of tensio-active agent.When its amount increases, the amount being equivalent to tensio-active agent increases, so the size of Nano microsphere can diminish.
Embodiment 3:
(1) 5mL vinylbenzene (analytical pure is got; through underpressure distillation except stopper) and 0.05g N; N; N-trimethyl-ethylene base benzene first ammonium chloride (VBTAC) joins in the three-necked bottle of the 500mL containing 185mL deionized water; mechanical stirring (400rpm) 30 minutes under room temperature, nitrogen protection; oxygen in removing reaction system, is then warmed up to 70 DEG C, adds 10mL, containing 0.37mmol azo-bis-isobutyrate hydrochloride (V
50) aqueous solution initiated polymerization of initiator, carry out 10h under being aggregated in the stirring velocity of nitrogen protection, 400rpm.Be polymerized the Nano microsphere that obtains under the rotating speed of 18500rpm centrifugal for 3 times, and with deionized water wash 3 times, remove unreacted, oligopolymer, initiator etc., be re-dispersed in 100mL deionized water, just obtained mass concentration is the Nano microsphere solution of 2.92% surface band positive charge.
(2) get 10mL Nano microsphere solution to add 10mL deionized water and add 3mL, 100 μ g/mL DSA again in three-necked bottle, put into stirrer, 50 DEG C of reaction 4h are heated under magnetic stirs, reacted rear fluorescent nanometer microsphere under the rotating speed of 18500rpm centrifugal for 3 times, and with deionized water wash 3 times, remove the fluorescence molecule of non-compound, and be re-dispersed in 23mL deionized water, obtain the fluorescent nanometer microsphere solution with aggregation inducing Fluorescence Increasing character that mass concentration is the surface band positive charge of 1.27%.
We find, its character is similar with the properties of samples of preparation in embodiment 1, but size becomes large, is about 110nm (see Figure 11).This is because polymerization single polymerization monomer N, N, N-trimethyl-ethylene base benzene first ammonium chloride (VBTAC) is amphiphilic monomer, serves as the effect of tensio-active agent.When its amount reduces, the amount being equivalent to tensio-active agent reduces, so the size of Nano microsphere can become large.
Embodiment 4:
(1) 5mL vinylbenzene (analytical pure is got; through underpressure distillation except stopper) and 0.04g N; N; N-trimethyl-ethylene base benzene first ammonium chloride (VBTAC) joins in the three-necked bottle of the 500mL containing 185mL deionized water; mechanical stirring (400rpm) 30 minutes under room temperature, nitrogen protection; oxygen in removing reaction system, is then warmed up to 70 DEG C, adds 10mL, containing 0.37mmol azo-bis-isobutyrate hydrochloride (V
50) aqueous solution initiated polymerization of initiator, carry out 10h under being aggregated in the stirring velocity of nitrogen protection, 400rpm.Be polymerized the Nano microsphere that obtains under the rotating speed of 18500rpm centrifugal for 3 times, and with deionized water wash 3 times, remove unreacted, oligopolymer, initiator etc., be re-dispersed in 100mL deionized water, just obtained mass concentration is the Nano microsphere solution of 2.92% surface band positive charge.
(2) get 10mL Nano microsphere solution to add 10mL deionized water and add 3mL, 100 μ g/mL DSA again in three-necked bottle, put into stirrer, 50 DEG C of reaction 4h are heated under magnetic stirs, reacted rear fluorescent nanometer microsphere under the rotating speed of 18500rpm centrifugal for 3 times, and with deionized water wash 3 times, remove the fluorescence molecule of non-compound, and be re-dispersed in 23mL deionized water, obtain the fluorescent nanometer microsphere solution with aggregation inducing Fluorescence Increasing character that mass concentration is the surface band positive charge of 1.27%.
We find, its character is similar with the properties of samples of preparation in embodiment 1, but size becomes large, is about 140nm (see Figure 12).This is because polymerization single polymerization monomer N, N, N-trimethyl-ethylene base benzene first ammonium chloride (VBTAC) is amphiphilic monomer, serves as the effect of tensio-active agent.When its amount reduces, the amount being equivalent to tensio-active agent reduces, so the size of Nano microsphere can become large.
Claims (4)
1. surface band positive charge has a fluorescent nanometer microsphere for aggregation inducing Fluorescence Increasing character, and it is prepared by following steps:
1) measure 5 ~ 10mL polymerization single polymerization monomer 1 to be scattered in the deionized water of 150 ~ 200mL, then add the polymerization single polymerization monomer 2 of 0.04g ~ 0.5g; Under room temperature, nitrogen protection, the oxygen in mechanical stirring removing reaction system; Add 10mL after being warming up to 70 ~ 90 DEG C, containing the aqueous solution of 0.3 ~ 0.5mmol initiator in reaction system, nitrogen protection, stir lower polyreaction 5 ~ 20 hours, obtain the Nano microsphere solution of surface band positive charge; After high speed centrifugation washing by the Nano microsphere that obtains again ultrasonic disperse in 100mL deionized water;
2) 10 ~ 20mL step 1 is measured) the Nano microsphere solution that obtains, add 10 ~ 20mL deionized water, and then add the AIE type fluorescence molecule of 3 ~ 5mL, 80 ~ 150 μ g/mL, 50 ~ 80 DEG C, under nitrogen protection condition, magnetic agitation reaction 4 ~ 10h; High speed centrifugation removes the AIE type fluorescence molecule of non-compound, obtains the fluorescent nanometer microsphere of the surface band positive charge of AIE molecule compound; The fluorescent nanometer microsphere obtained is scattered in deionized water, thus obtains the fluorescent nanometer microsphere with aggregation inducing Fluorescence Increasing character of surface band positive charge;
Polymerization single polymerization monomer 1 is vinylbenzene, fluorostyrene, methyl methacrylate or glycidyl methacrylate; Polymerization single polymerization monomer 2 is N, N, N-trimethyl-ethylene base benzene first ammonium chloride, 2-(diisopropylaminoethyl) β-dimethyl-aminoethylmethacrylate or 2-aminoethyl methacrylate hydrochloride.
2. a kind of surface band positive charge as claimed in claim 1 has the fluorescent nanometer microsphere of aggregation inducing Fluorescence Increasing character, it is characterized in that: initiator is azo-bis-isobutyrate hydrochloride, azo two isobutyl imidazoline hydrochloride or azo isobutyl cyano group methane amide.
3. a kind of surface band positive charge as claimed in claim 1 has the fluorescent nanometer microsphere of aggregation inducing Fluorescence Increasing character, it is characterized in that: the fluorescence molecule of AIE type is 9,10-bis-(styryl) anthracene sulfonic acid salt, 1,2-bis-[4-(3-sulfonic acid propoxy-) phenyl]-1,2-diphenylethlene disodium salt or 1,1,2,2-tetra-[4-(3-sulfonic acid propoxy-) phenyl] ethene tetra-na salt.
4. a kind of surface band positive charge of claims 1 to 3 described in any one has the application of fluorescent nanometer microsphere in bio-imaging of aggregation inducing Fluorescence Increasing character.
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