CN101701151A - Fluorescent nanoparticle and preparation method and application thereof - Google Patents

Fluorescent nanoparticle and preparation method and application thereof Download PDF

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CN101701151A
CN101701151A CN 200910203407 CN200910203407A CN101701151A CN 101701151 A CN101701151 A CN 101701151A CN 200910203407 CN200910203407 CN 200910203407 CN 200910203407 A CN200910203407 A CN 200910203407A CN 101701151 A CN101701151 A CN 101701151A
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nano particles
fluorescent nano
colloidal sol
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rare earth
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CN101701151B (en
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王远
符小艺
邵光胜
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Peking University
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Abstract

The invention discloses a class of fluorescent nanoparticles and a preparation method and an application thereof. The fluorescent nanoparticle comprises matrix material and fluorescent dye dispersed in the matrix material, wherein the matrix material is the compound composed of hydrophobic polymer main chain of which is hydrocarbon chain, organo-siloxane and organosilicon-polymer, and the fluorescent dye is rare earth complex with photoluminescence performance. The preparation method of the fluorescent nanoparticle comprises the following steps: dissolving rare earth complex, hydrophobic polymer and compound RSi (OR') 3 into organic solvent miscible with water; adding the mixture into aqueous solution containing surfactant; using precipitation and hydrolytic condensation reaction to form nanoparticles. The nanoparticle of the invention has favourable luminous performance and favourable stability, can have a silicon oxide shell and a surface functional group and can be used for bonding biomolecules; biological probes based on the fluorescent nanoparticle have wide application prospects on the aspects of high-sensitivity fluorescence immunoassay, imaging and the like.

Description

Fluorescent nano particles and its production and application
Technical field
Fluorescent nano particles of the present invention and its production and application, particularly a class is based on the fluorescent nano particles of rare earth compounding fluorescence dye and preparation method thereof and application.
Background technology
Fluorescent probe based on luminescent rare earth complexes has the important application prospect at aspects such as bio-sensing, cell and imaging of tissue.Its major cause is that rare earth ion can reduce the fluorescence interference that biomaterial itself produces in the sharp line emission and the long luminous attitude life-span of visible or near-infrared region, be expected to make to detect to have very high sensitivity and efficient, avoided a series of problems of using radioelement to bring simultaneously.
Because the f-f transition of rare earth ion is subjected to eelctric dipole to select the restriction of rate, the light absorpting ability of rare earth ion itself is very weak, molar extinction coefficient is very little, therefore to obtain bigger photoluminescence efficiency, need carry out sensitization to it, promptly pass through part (or charge transfer state) extinction, and make energy be transferred to the rare earth luminescence attitude to realize the efficiently luminous of rare earth ion.Many rare earth compoundings can be obtained luminous intensity preferably by ultraviolet excitation.Yet uv excitation light is bigger to organism damage and interference, and the UV-light penetration depth is little, can produce stronger background fluorescence simultaneously, makes the application of ultraviolet excited fluorescence probe be subjected to a lot of restrictions.Therefore, to the window that excites of long wave direction expansion rare earth compounding, the rare earth coordination compound fluorescent probe that acquisition has good visible light or near infrared light stimulated luminescence ability becomes one of important development direction of rare earth compounding biological probe.Rare earth compounding with good visible light and near infrared light stimulated luminescence performance is very limited.
In recent years, the present inventor designs, has synthesized europium complex [Eu (tta) 3Dpbt] (tta is the thenoyltrifluoroacetone negative ion for dpbt=2-(N, N-Diethyl Aniline-4-yl)-4,6-two (3-1-yl)-1,3,5-triazines) (Y.Wang, et al., Angew.Chem.Int.Ed.2004,43,5010).On this basis, we design, have synthesized a series of novel rare-earth title complexs, and they all have good visible light and near infrared light stimulated luminescence performance (Y.Wang, et al., Adv.Funct.Mater.2007,17,3663; Wang Yuan, Hao Rui has the europium complex and the synthetic method thereof of excited by visible light luminescent properties, CN 101066974A).If with this type of rare earth compounding serves as that basis preparation biological fluorescent labeling is applied to bioanalysis, the employing longwave optical excites, will make bioanalysis have highly sensitive, high s/n ratio, penetration depth big, to the organism damage with disturb advantages such as little, that analysis cost is low.Yet known rare earth compounding with visible and the efficient stimulated luminescence performance of near infrared light is a hydrophobic molecule, and its stability in the aqueous solution is not high, thereby is difficult to directly use in the aqueous solution these complex molecules as biological fluorescent labeling.In addition, even can by ultraviolet excitation efficient luminous be the rare earth compounding of part with multiple tooth carboxyl, because the ligand exchange reaction that may exist between rare earth compounding and the biomacromolecule also makes it be very limited in the application aspect biological detection and the imaging.
One that solves this type of problem is that rare earth compounding is embedded in fluoropolymer resin or the inorganic nano-particle than feasible program.West roc (western roc, Gu Xiaohua, Huang Xiangan, He Yuxian, inorganic-organic core-shell type rare earth polymer materials and preparation method thereof, CN1966534; ) wait the hydrophobic rare earth compounding that adopts emulsion polymerization and swelling entrapping method will have the ultraviolet excitation luminescent properties to be embedded in the fluoropolymer resin, prepared nanoparticle with ultraviolet excitation luminescent properties; (Chen Yang is based on fluorescence energy transfer principle luminous rare-earth nanometer particles and preparation method, CN 1304523C for Chen Yang and Yuan Jingli etc.; Yuan Jingli, Tan Mingqian, Ye Zhiqiang, Wang Guilan, a kind of functional nano rare earth particulate and preparation and application, CN1298807C) adopt respectively microemulsion method or Method is embedded in hydrophilic rare earth compounding in the nanoparticle that silicon-dioxide is matrix, has prepared the rare earth compounding based nano-material with ultraviolet excitation luminescent properties.The nanoparticle that common hydrophobic polymer is a matrix dispersed undesirable in the aqueous solution easily reunited, and in many organic solvents because of the polymer swelling, cause easily that fluorescence molecule leaks in the nanoparticle.The wetting property of silica matrix and water-insoluble rare earth compounding is relatively poor, when the silicon oxide that forms with hydrolysis reaction wraps up water-insoluble rare earth compounding synthesizing nano-particle, usually cause the stability of the fluorescent nano particles that forms not high owing to be difficult to form fine and close integument.In addition, when the silicon oxide or derivatives thereof that forms with hydrolysis reaction wraps up rare earth compounding, often exist between Si-OH and the fluorescence molecule to interact, thereby the fluorescence quantum yield of reduction rare earth compounding weakens luminous intensity.(Chem.Comm.2008 is 365-367) to inventor's design, synthetic rare earth compounding [Eu (tta) for Jingli Yuan, et.al. for Yuan Jingli etc. 3Dpbt] carried out the structure modulation, to contain-Si (OR ') 3The CDHH of group replaces [Eu (tta) 3Dpbt] in assistant ligand tta, utilize prepared with microemulsion reactor a kind of can be by the luminous rare earth compounding base fluorescent nano particles of excited by visible light, though it excites the window afterbody to extend to visible region, but its excitation peak orchid moves to the ultraviolet region, the optical extinction coefficient of visible region is little, about 0.2 (excitation wavelength is 335nm) of fluorescence quantum yield, show that its excited by visible light luminous power a little less than.
Ideal rare earth compounding base namo fluorescence probe should have following feature: 1) have good dispersiveness and stable in the aqueous solution; 2) rare earth compounding in the nanoparticle or its aggregate have strong photoluminescence ability, particularly visible light and near infrared light stimulated luminescence ability; 3) nanoparticle surface has the active group that can supply with the biomolecules bonding.
The inventor once adopted the rare earth compounding base fluorescent nano particles synthetic method parcel that has been seen in report is designed, synthetic has excellent visible light and near infrared light stimulated luminescence performance rare earth compounding with synthesizing new rare earth compounding base fluorescent nano particles, for example micro emulsion polymerization, RSi (OR ') 3Hydrolysis entrapping method etc., but all be difficult to obtain gratifying result.Its one of the main reasons be have the part of rare earth compounding of good visible light and near infrared stimulated luminescence performance and a rare earth ion coordination a little less than, common title complex embedding method causes it that ligand exchange reaction takes place easily, loses original stimulated luminescence characteristic.
Summary of the invention
The purpose of this invention is to provide new fluorescent nano particles based on the rare earth compounding fluorescence dye and preparation method thereof and application.
Fluorescent nano particles provided by the present invention comprises substrate material and the fluorescence dye that is dispersed in the described substrate material; Described substrate material is for being the mixture that hydrophobic polymer, organo-siloxane and the organosilicon polymer of hydrocarbon chain forms by main chain; Described fluorescence dye is the rare earth compounding with photoluminescence performance.Main chain is that hydrophobic polymer, the organo-siloxane of hydrocarbon chain, the mol ratio of rare earth compounding are 0.1～10: 0.005～3 in the described fluorescent nano particles: 1, and the quality percentage composition of organosilicon polymer is 0.5%～40% in the described fluorescent nano particles.
Above-mentioned main chain is that the hydrophobic polymer of hydrocarbon chain can be one or more the miscellany of arbitrary combination in polymethacrylate, polyacrylic ester, polystyrene, methacrylic ester styrol copolymer, acrylate styrol copolymer, polymaleic acid dialkyl, dialkyl maleate styrol copolymer and the MBS.
Above-mentioned organo-siloxane is RSi (OR ') 3Wherein R is the alkyl that 3～18 alkyl, thiazolinyl, phenyl or fluorine replace for the C carbonatoms, and R ' is 1～2 alkyl for the C carbonatoms.Above-mentioned organosilicon polymer is described organosilicone compounds RSi (OR ') 3Hydrolysis and the netted organosilicon polymer that forms by the condensation of silicon hydroxyl.
Above-mentioned nanoparticle surface contains also that have can be for the active group of graft reaction.Described active function groups for graft reaction is hydroxyl, amino, carboxyl or sulfydryl.
The skin of above-mentioned nanoparticle also can be enclosed with the silicon oxide shell.Described silicon oxide shell thickness is 1～20nm.
Above-mentioned surface with fluorescent nano particles of silicon oxide shell can contain also that have can be for the active group of graft reaction, and described active function groups for graft reaction is hydroxyl, amino, carboxyl or sulfydryl.
The particle diameter of above-mentioned nanoparticle is 10-200nm.
The above-mentioned rare earth compounding that is scattered in the nanoparticle-based material is the rare earth compounding with under visible light and/or near infrared light and/or ultraviolet excitation visible emitting or near infrared light performance; Described rare earth compounding comprises europium complex and/or terbium coordination compound and/or ytterbium title complex and/or erbium title complex and/or neodymium title complex.Described europium complex can be structure suc as formula the compound shown in I or the formula II.
Figure G2009102034073D00031
Formula I formula II
Among above-mentioned formula I and the formula II, R 1, R 2Be that carbonatoms is 1 to 4 alkyl, R 3, R 4, R 5, R 6Be methyl or H, R 7, R 8Be H, methyl or three fluoro methyl;
The preparation method of above-mentioned fluorescent nano particles provided by the present invention comprises the steps:
1) with rare earth compounding, main chain is the hydrophobic polymer of hydrocarbon chain and compound R Si (OR ') 3Be dissolved in can with the miscible organic solvent of water in, make that to contain concentration be 1.0 * 10 -6～1.0 * 10 -3The above-mentioned rare earth compounding of mol/L, concentration are 1.0 * 10 -6～1.0 * 10 -2The main chain of mol/L is the hydrophobic polymer of hydrocarbon chain; Concentration is 1.0 * 10 -6～1.0 * 10 -2The compound R Si of mol/L (OR ') 3Solution;
2) under stirring or ultrasound condition, the solution that step 1) is obtained joins volume in its aqueous solution that contains dispersion agent of 1～100 times, obtain with the main chain is that the hydrophobic polymer, organo-siloxane of hydrocarbon chain and mixture that organosilicon polymer forms are substrate material, colloidal sol based on the fluorescent nano particles of rare earth compounding is designated as colloidal sol C with it.When dispersion agent is tensio-active agent or during water soluble organic polymer, its concentration of aqueous solution 1.0 * 10 -4～1.0 * 10 -2Mol/L; When dispersion agent was protein, its concentration of aqueous solution was 0.1～5.0mg/mL.
Described can be one or more any mixing in methyl alcohol, ethanol, acetone, acetonitrile, dimethyl formamide and the tetrahydrofuran (THF) with the miscible organic solvent of water; Described dispersion agent comprises tensio-active agent, water soluble protein or water soluble organic polymer; That described tensio-active agent comprises is cationic, anionic, non-ionic type or amphoterics or their mixture; Described protein comprises serum albumin or ovalbumin etc.; Described water soluble organic polymer comprises Polyvinylpyrolidone (PVP) (PVP), polyvinyl alcohol (PVA) etc.; Described main chain is that the hydrophobic polymer of hydrocarbon chain is one or more the miscellany of arbitrary combination in polymethacrylate, polyacrylic ester, polystyrene, methacrylic ester styrol copolymer, acrylate styrol copolymer, polymaleic acid dialkyl, dialkyl maleate styrol copolymer and the MBS, RSi (OR ") 3In R be the alkyl that 3～18 alkyl, thiazolinyl, phenyl or fluorine replace for the C carbonatoms, R " be 1～2 alkyl for the C carbonatoms.
Above-mentioned steps 2) in, it is 1.0 * 10 that described aqueous dispersant can be concentration -4～1.0 * 10 -3The water phase surfactant mixture of mol/L, the volume ratio of the solution that described water phase surfactant mixture and described step 1) obtain can be 1-50: 1.
In the described method, can comprise that also with concentration be 1.0 * 10 -6～1.0 * 10 -4The XRSi of mol/L (OR ") 3Alcoholic solution join described step 2) in the colloidal sol (colloidal sol C) of the fluorescent nano particles that obtains, make described RSi (OR ') 3With described XRSi (OR ") 3The usage quantity mol ratio be 1-200: 1, in 0～80 ℃ of reaction 1～24hr, making described surface contains and can be used for the active function groups of other compound covalent bonding, be that mixture that hydrophobic polymer, organo-siloxane and the organosilicon polymer of hydrocarbon chain forms is a substrate material with the main chain, colloidal sol based on the fluorescent nano particles of rare earth compounding fluorescence dye is designated as colloidal sol E with it; Described compounds X RSi (OR ") 3In, X is amino, NH 2CH 2CH 2NH-base, carboxyl or sulfydryl, XR be carbonatoms be 2～5 contain the substituent alkyl of X, OR " be methoxyl group, oxyethyl group, OH or ONa.
Aforesaid method can be included in also in the described solution of step 1) that to add final concentration be 1.0 * 10 -6～1.0 * 10 -4The XRSi of mol/L (OR ") 3And in step 2) described this solution is added after the described aqueous dispersant, in 0～80 ℃ of reaction 1～24hr, making described surface contains and can be used for the active function groups of other compound covalent bonding, be that mixture that hydrophobic polymer, organo-siloxane and the organosilicon polymer of hydrocarbon chain forms is a substrate material with the main chain, colloidal sol based on the fluorescent nano particles of rare earth compounding fluorescence dye is designated as colloidal sol G with it; Described compounds X RSi (OR ") 3In, X is amino or sulfydryl, XR be carbonatoms be 2～5 contain the substituent alkyl of X, OR " be methoxyl group, oxyethyl group.
In the described method, comprise that also with concentration be 5.0 * 10 -3Mol/L～5.0 * 10 -1The positive silicon ester alcoholic solution of mol/L joins in the colloidal sol (colloidal sol C, colloidal sol E or colloidal sol G) of above-mentioned fluorescent nano particles according to 1: 5～100 volume ratio, in 0～80 ℃ of reaction 1～24hr, centrifugation, precipitation is scattered in the water, makes the colloidal sol of fluorescent nano particles with shell-core structure; Then, add 1.0 * 10 -6～1.0 * 10 -4The XRSi of mol/L (OR ") 3Alcoholic solution, in 0～80 ℃ of reaction 1～24hr, make have shell-core structure, the surface contains the colloidal sol that can be used for the fluorescent nano particles of the active function groups of other compound covalent bonding; Wherein, the alcoholic solution of described positive silicon ester is the methyl alcohol or the ethanolic soln of methyl silicate or tetraethoxy.Described compounds X RSi (OR ") 3In, X is amino, NH 2CH 2CH 2NH-base, carboxyl or sulfydryl, XR be carbonatoms be 2～5 contain the substituent alkyl of X, OR " be methoxyl group, oxyethyl group, OH or ONa.
In the aforesaid method, described main chain be hydrocarbon chain, compound R Si (OR ') 3And the mol ratio of rare earth compounding usage quantity is 0.1～10: 0.1～10: 1.
In the described method, comprise that also the fluorescent nano particles to obtaining carries out purifying, the method for described purifying will be deposited in then and disperse in the water for the centrifugal and collecting precipitation of colloidal sol of the fluorescent nano particles that will obtain, and perhaps carry out purifying with chromatography.
Fluorescent nano particles of the present invention has following advantage:
1, has excellent luminescent properties.Fluorescent nano particles based on rare earth compounding has the advantage that emission peak is narrow, intensity big, the Stocks displacement is big.When adopting the rare earth compounding with visible and near infrared stimulated luminescence of the present invention to be fluorescence dye, prepared fluorescent nano particles excites to descend to have very high rare-earth fluorescent quantum yield at visible light and near infrared light, and is for example of the present invention many based on [Eu (tta) 3Dpbt] and [Eu (fod) 3Dpbt] the fluorescence quantum yield of fluorescent nano particles under excited by visible light greater than 0.3, its excited by visible light luminosity is far above the rare earth compounding base nanoparticle luminosity under the same conditions of other document and patent report.The rare earth compounding of the same race that rare earth compounding base fluorescent nano particles more of the present invention are dissolved in the solvent has better long wave excited by visible light luminescent properties, for example, and with [the Eu (tta) that is dissolved in the toluene 3Dpbt] compare, of the present invention based on [Eu (tta) 3Dpbt] fluorescent nano particles the visible region excitation peak and excite the red limit of window that remarkable red shift takes place, this excitation peak that causes by the rare earth compounding aggregate and excite the phenomenon of the window remarkable red shift in red limit not appear in the newspapers as yet.Above-mentioned fluorescent nano particles has excellent two-photon excitation luminescent properties simultaneously, excites down at near-infrared laser, can send bright ruddiness.Have an excellent two-photon excitation luminescent properties and can satisfy synthesis of nano biological probe needs, do not appear in the newspapers as yet based on the fluorescent nano particles of rare earth compounding fluorescence dye.In addition, the invention solves the nanometer embedding problem of the hydrophobic rare earth compounding with near infrared light emitting performance, the nanoparticle that institute's synthetic is surrounded by this type of rare earth compounding fluorescence dye has good near infrared light emitting performance.Above-mentioned excellent photoluminescence performance make rare earth compounding base fluorescent nano particles of the present invention be highly suitable for synthetic highly sensitive, penetration depth big, to the little nano fluorescent bioprobe of biological sample damage.
2, stability is good.In fluorescent nano particles of the present invention, rare earth compounding is scattered in the mixture substrate material of hydrophobic polymer, organo-siloxane and organosilicon polymer formation that main chain is a hydrocarbon chain, because polymkeric substance good hydrophobic performance, ion in the water molecules and the aqueous solution is difficult to see through polymer layer and title complex effect, and the rare earth compounding in the substrate material also is difficult to let out.Above-mentioned special construction makes luminescent nanoparticle of the present invention have good stable luminescent property.On the other hand, the outer field silicon hydroxyl of fluorescent nano particles of the present invention has good hydrophilicity, makes fluorescent nano particles of the present invention have good dispersiveness in the aqueous solution, and its colloidal solution has good dispersion stabilization.And the use of tensio-active agent has further improved its dispersion stabilization, and colloidal solution can not produce precipitation through long-term placement.
3, preparation method's controllability of the present invention is strong, and the functional group on fluorescent nano particles surface can be used for the bonding of biomolecules.
The preparation method of fluorescent nano particles of the present invention is the co-precipitation pack, be about to rare earth compounding, main chain and be hydrocarbon chain with compound R Si (OR ') 3Be dissolved in can be miscible with water organic solvent, its adding is contained in the aqueous solution of described dispersion agent, utilize precipitation and hydrolysis-condensation reaction, form the nanoparticle of the described matrix that is surrounded by the rare earth compounding fluorescence dye; The present invention provides simultaneously has the silicon oxide shell and the surface has the described fluorescent nano particles and a synthetic method thereof of active group.Change the rare earth compounding among the present invention into other fluorescence dye, also can prepare the fluorescent nano particles of other fluorescence dye by method provided by the invention.Siloxanes, three's ratio that main chain is hydrocarbon chain and organic solvent and dispersion agent type by modulation rare earth compounding, band hydrophobic chain can easily be regulated and control parameters such as fluorescent nano particles middle-weight rare earths title complex content, particle sizes.And the kind of fluorescent nano particles surfactivity functional group and content can be controlled by selecting siloxanes kind and add-on thereof.
Fluorescent nano particles of the present invention dispersion stabilization in the aqueous solution is good, and the surface has an active function groups that can be used for the biomolecules covalent bonding, therefore can be directly used in the mark of biomolecules.Various albumen as immunoglobulin IgG and anti-IgG, all can adopt fluorescent nano particles provided by the present invention to carry out mark.The nano fluorescent bioprobe that obtains behind the mark can be used for highly sensitive fluoroimmunoassay, fields such as bioluminescence imaging and biochip.
Description of drawings
Fig. 1 is based on [Eu (tta) 3Dpbt] transmission electron microscope photo of fluorescent nano particles of title complex; A is the transmission electron microscope photo (nano-particles size scope 10～50nm, median size 27nm) of the fluorescent nano particles of embodiment 11 preparations among Fig. 1; B is the transmission electron microscope photo (size range 50～90nm, median size 74nm) of the fluorescent nano particles of embodiment 1 preparation.
Fig. 2 be embodiment 1 preparation based on [Eu (tta) 3Dpbt] the excitation spectrum (λ of fluorescent nano particles Em=614nm).
Fig. 3 be embodiment 1 preparation based on [Eu (tta) 3Dpbt] the emission spectrum (λ of fluorescent nano particles Ex=412nm).
Fig. 4 is [Eu (tta) in fluorescent nano particles and the nanometer particle colloid that obtains behind the anti-human IgG covalent bonding of organic fluorescent dye FITC labelled goat 3Dpbt] excitation spectrum (A) (λ Em=614nm) with emission spectrum (B) (λ Ex=412nm) and the excitation spectrum of FITC (A) (λ Em=530nm) with emission spectrum (B) (λ Ex=470nm).
Fig. 5 be among the embodiment 29 fluorescence intensity with human IgG change in concentration figure.
Embodiment
In order to be described more specifically the present invention, now provide some embodiment, but content involved in the present invention is not limited only to these embodiment.Method therefor is ordinary method if no special instructions among the following embodiment.
Among the following embodiment, tta is the thenoyltrifluoroacetone negative ion, and fod is 6,6,7,7,8,8,8-seven fluoro-2,2-dimethyl octane-3,5-diketone negative ion.
Embodiment 1: with the mixture of PMMA, organo-siloxane and organosilicon polymer be matrix based on [Eu (tta) 3Dpbt] the preparation of fluorescent nano particles
With octyl group Trimethoxy silane (OTS), polymethylmethacrylate (PMMA) and [Eu (tta) 3Dpbt] (its structural formula is shown in formula III; Y.Wang, et al., Angew.Chem.Int.Ed.2004,43,5010) be dissolved in the acetone, being mixed with OTS concentration is 7.6 * 10 -4Mol/L, PMMA concentration is 1.6 * 10 -5Mol/L, [Eu (tta) 3Dpbt] concentration is 1.6 * 10 -4The acetone soln of mol/L is designated as solution A 1.Under room temperature and agitation condition, it is 1.0 * 10 that 25mL solution A 1 is added dropwise to 75mL concentration -3In the aqueous solution (being designated as solution B 1) of the hexadecyl trimethoxy brometo de amonio (CTAB) of mol/L (solution A 1 is 1: 3 with solution B 1 volume ratio), stir 10min, obtain mixture with PMMA, organo-siloxane and organosilicon polymer (OTS hydrolysis and the network polymer by silicon hydroxyl condensation formation) and be matrix, be surrounded by [Eu (tta) 3Dpbt] the colloidal sol (being designated as colloidal sol C1) of fluorescent nano particles.
With 1mL concentration is 3.7 * 10 -53-TSL 8330 (APS) the methanol solution D1 of mol/L adds among the 100mL colloidal sol C1, in 60 ℃ of reactions 1 hour, centrifugation, the gained precipitation is scattered in the pure water again, makes the colloidal sol (being designated as colloidal sol E1) that the surface is connected to the fluorescent nano particles of the amino group that can supply graft reaction.
Transmission electron microscope (TEM) test result shows that the median size of fluorescent nano particles is 74nm among the colloidal sol C1 of above-mentioned preparation.The transmission electron microscope photo of the fluorescent nano particles among the colloidal sol E1 of above-mentioned preparation is shown in b among Fig. 1, and the distribution of sizes scope of obtained fluorescent nano particles is at 50～90nm, median size 74nm.Because the total mass of fluorescent nano particles among the add-on little colloidal sol C1 far away of APS, thus Electronic Speculum to characterize the size of nanoparticle in C1 colloidal sol that experiment records and the E1 colloidal sol very approaching.
With prepared fluorescent nano particles centrifugation, with water washing, drying, with the OTS in its medium of acetone extract, gas-chromatography and icp analysis result show, organo-siloxane OTS and Eu mol ratio are 0.5 in the nanoparticle, hydrophobic polymer PMMA and Eu ionic mol ratio are 0.1 in the fluorescent nano particles, and the quality percentage composition of organosilicon polymer (OTS hydrolysis and the network polymer that forms by the condensation of silicon hydroxyl) is 32%.
The fluorescence spectrum test result shows that fluorescent nano particles has excellent visible light stimulated luminescence performance among the colloidal sol C1 of above-mentioned preparation, and its excitation peak in the visible region is positioned at 412nm, and afterbody is extended to 480nm.Fluorescence excitation spectrum (the λ of the fluorescent nano particles colloidal sol E1 of above-mentioned preparation Em=614nm) as shown in Figure 2, the fluorescence emission spectrum (λ of the fluorescent nano particles colloidal sol E1 of above-mentioned preparation Em=412nm) as shown in Figure 3.The result shows that above-mentioned fluorescent nano particles has excellent visible light stimulated luminescence performance, and its excitation peak in the visible region is positioned at 412nm, and afterbody is extended to 480nm.Propanol solution with 4-dicyanomethylene-2-methyl-6-p-dimethylaminostyrl-4H-py ran (DCM) is reference substance (Φ=0.57), at wavelength is under the 410nm excitation light irradiation, and the europium ion fluorescence quantum yield that records this fluorescent nano particles is 0.30.At wavelength is that the near-infrared laser of 832 nanometers excites down, and above-mentioned fluorescent nano particles can send bright ruddiness, is that to record its two-photon excitation action section be 40GM (1GM=10 to reference substance with Rhodamine B -50Cm 4S photo -1Molecule -1).
(formula III)
Embodiment 2: with the mixture of styrene-methyl methacrylate copolymer, organo-siloxane and organosilicon polymer be matrix based on [Eu (tta) 3Dpbt] the preparation of fluorescent nano particles
According to the method among the embodiment 1, substitute PMMA preparation A2 with styrene-methyl methacrylate copolymer (P (St-co-MMA)), each solute concentration is in this solution: OTS, 7.6 * 10 -4Mol/L; P (St-co-MMA), 1.6 * 10 -4Mol/L; [Eu (tta) 3Dpbt], 1.6 * 10 -4Mol/L; Under room temperature and agitation condition, it is 1.4 * 10 that 30mL solution A 2 is added dropwise to 70mL concentration -3Stir 10min in the aqueous solution of the CTAB of mol/L (being designated as solution B 2), obtain mixture with P (St-co-MMA), organo-siloxane and organosilicon polymer (OTS hydrolysis and the network polymer by silicon hydroxyl condensation formation) and be matrix, be surrounded by [Eu (tta) 3Dpbt] the colloidal sol (being designated as colloidal sol C2) of fluorescent nano particles.
The TEM test result shows that the distribution of sizes scope of fluorescent nano particles is at 22～85nm among the colloidal sol C1 of above-mentioned preparation, and median size is 45nm.With prepared fluorescent nano particles centrifugation, with water washing, drying, with the OTS in its medium of acetone extract, gas-chromatography and icp analysis result show, the mol ratio of organo-siloxane OTS and Eu is 1.1 in the nanoparticle, hydrophobic polymer P (St-co-MMA) is 1 with Eu ionic mol ratio in the nanoparticle, and the mass percent content of organosilicon polymer is 32%.
The fluorescence spectrum test result shows that fluorescent nano particles has excellent visible light stimulated luminescence performance among the colloidal sol C 1 of above-mentioned preparation, and its excitation peak in the visible region is positioned at 415nm, and afterbody is extended to 475nm.Propanol solution with DCM is reference substance (Φ=0.57), is under the 415nm excitation light irradiation at wavelength, and the europium ion fluorescence quantum yield that records this fluorescent nano particles is 0.31.At wavelength is that the near-infrared laser of 830 nanometers excites down, and above-mentioned fluorescent nano particles can send bright ruddiness, is that to record its two-photon excitation action section be 50GM (1GM=10 to reference substance with Rhodamine B -50Cm 4S photo -1Molecule -1).
Embodiment 3: with the mixture of PMMA, organo-siloxane and organosilicon polymer be matrix based on [Eu (tta) 3Dpbt] the preparation of fluorescent nano particles
According to the method among the embodiment 2, substitute P (St-co-MMA) with PMMA, acetonitrile substitutes acetone preparation A3; Substitute CTAB obtain solution B3 with bovine serum albumin, the concentration of bovine serum albumin is 3.0mg/mL.According to preparation method among the embodiment 2 and condition, prepare mixture with PMMA, OTS and organosilicon polymer (OTS hydrolysis and by silicon hydroxyl condensation formation) and be substrate material, based on [Eu (tta) 3Dpbt] the colloidal sol of fluorescent nano particles, be designated as colloidal sol C3.
The TEM test result shows that the distribution of sizes of fluorescent nano particles is 10～40nm among the colloidal sol C3 of above-mentioned preparation, and median size is 25nm.With prepared fluorescent nano particles centrifugation, with water washing, drying, with the OTS in its medium of acetone extract, gas-chromatography and icp analysis result show, the mol ratio of organo-siloxane OTS and Eu is 0.1 in the nanoparticle, hydrophobic polymer PMMA and Eu ionic mol ratio are 1 in the nanoparticle, and the mass percent content of organosilicon polymer is 5%.
The fluorescence spectrum test result shows that fluorescent nano particles has excellent visible light stimulated luminescence performance among the colloidal sol C3 of above-mentioned preparation, and its excitation peak in the visible region is positioned at 412nm, and afterbody is extended to 475nm.At wavelength is that the near-infrared laser of 830 nanometers excites down, and above-mentioned fluorescent nano particles can send bright ruddiness.
Embodiment 4: with the mixture of styrene-methyl methacrylate copolymer, organo-siloxane and organosilicon polymer be matrix based on [Eu (tta) 3Dpbt] the preparation of fluorescent nano particles
According to the method among the embodiment 2, substitute PMMA with P (St-co-MMA), tetrahydrofuran (THF) substitutes acetone preparation A4; Human serum albumin substitutes CTAB obtain solution B4, and the concentration of human serum albumin is 5.0mg/ml.According to preparation method among the embodiment 1 and condition, prepare mixture with P (St-co-MMA), OTS and organosilicon polymer (OTS hydrolysis and by silicon hydroxyl condensation formation) and be substrate material, based on [Eu (tta) 3Dpbt] the colloidal sol of fluorescent nano particles, be designated as colloidal sol C4.
The TEM test result shows that the distribution of sizes of fluorescent nano particles is 40～100nm among the colloidal sol C4 of above-mentioned preparation, and median size is 60nm.With prepared fluorescent nano particles centrifugation, with water washing, drying, with the OTS in its medium of acetone extract, gas-chromatography and icp analysis result show, the mol ratio of organo-siloxane OTS and Eu is 1.0 in the nanoparticle, hydrophobic polymer P (St-co-MMA) is 1 with Eu ionic mol ratio in the nanoparticle, and the mass percent content of organosilicon polymer is 30%.
The fluorescence spectrum test result shows that fluorescent nano particles has excellent visible light stimulated luminescence performance among the colloidal sol C4 of above-mentioned preparation, and its excitation peak in the visible region is positioned at 412nm, and afterbody is extended to 475nm.At wavelength is that the near-infrared laser of 830 nanometers excites down, and above-mentioned fluorescent nano particles can send bright ruddiness.
Embodiment 5: with the mixture of polystyrene, organo-siloxane and organosilicon polymer be matrix based on [Eu (tta) 3Dpbt] the preparation of fluorescent nano particles
According to the method among the embodiment 2, substitute PMMA with polystyrene (PST), dimethyl formamide substitutes acetone preparation A5; Substitute CTAB obtain solution B5 with chicken egg white, the concentration of chicken egg white is 0.5mg/mL.According to preparation method among the embodiment 1 and condition, prepare mixture with PST, OTS and organosilicon polymer (OTS hydrolysis and by silicon hydroxyl condensation formation) and be substrate material, based on [Eu (tta) 3Dpbt] the colloidal sol of fluorescent nano particles, be designated as colloidal sol C5.
The TEM test result shows that the distribution of sizes of fluorescent nano particles is 100～200nm among the colloidal sol C5 of above-mentioned preparation, and median size is 160nm.With prepared fluorescent nano particles centrifugation, with water washing, drying, with the OTS in its medium of acetone extract, gas-chromatography and icp analysis result show, the mol ratio of organo-siloxane OTS and Eu is 0.6 in the nanoparticle, hydrophobic polymer PST and Eu ionic mol ratio are 0.1 in the nanoparticle, and the mass percent content of organosilicon polymer is 25%.
The fluorescence spectrum test result shows that fluorescent nano particles has excellent visible light stimulated luminescence performance among the colloidal sol C5 of above-mentioned preparation, and its excitation peak in the visible region is positioned at 412nm, and afterbody is extended to 475nm.At wavelength is that the near-infrared laser of 830 nanometers excites down, and above-mentioned fluorescent nano particles can send bright ruddiness.
Embodiment 6: with the mixture of styrene-methyl methacrylate copolymer, organo-siloxane and organosilicon polymer be matrix based on [Eu (tta) 3Dpbt] the preparation of fluorescent nano particles
According to the method among the embodiment 2, substitute OTS preparation A2 with octadecyltriethoxy silane, each solute concentration is in this solution: octadecyltriethoxy silane, 1.6 * 10 -3Mol/L; P (St-co-MMA), 1.6 * 10 -3Mol/L; [Eu (tta) 3Dpbt], 1.6 * 10 -4Mol/L; Under room temperature and agitation condition, it is 1.0 * 10 that 30mL solution A 2 is added dropwise to 70mL concentration -3Stir 10min in the water-soluble polymer PVP aqueous solution (being designated as solution B 2) of mol/L, obtain mixture with P (St-co-MMA), organo-siloxane and organosilicon polymer (hydrolysis and the network polymer by silicon hydroxyl condensation formation) and be matrix, be surrounded by [Eu (tta) 3Dpbt] the colloidal sol (being designated as colloidal sol C2) of fluorescent nano particles.
The TEM test result shows that the distribution of sizes scope of fluorescent nano particles is at 60～150nm among the colloidal sol C1 of above-mentioned preparation, and median size is 100nm.With prepared fluorescent nano particles centrifugation, with water washing, drying, with the octadecyltriethoxy silane in its medium of acetone extract, gas-chromatography and icp analysis result show, the mol ratio of octadecyltriethoxy silane and Eu is 3 in the nanoparticle, hydrophobic polymer P (St-co-MMA) is 10 with Eu ionic mol ratio in the nanoparticle, and the quality percentage composition of organosilicon polymer (octadecyltriethoxy silane hydrolysis and the network polymer that forms by the condensation of silicon hydroxyl) is 35%.
The fluorescence spectrum test result shows that fluorescent nano particles has excellent visible light stimulated luminescence performance among the colloidal sol C1 of above-mentioned preparation, and its excitation peak in the visible region is positioned at 415nm, and afterbody is extended to 475nm.Propanol solution with DCM is reference substance (Φ=0.57), is under the 415nm excitation light irradiation at wavelength, and the europium ion fluorescence quantum yield that records this fluorescent nano particles is 0.31.At wavelength is that the near-infrared laser of 830 nanometers excites down, and above-mentioned fluorescent nano particles can send bright ruddiness.
Embodiment 7: with the mixture of PMMA, organo-siloxane and organosilicon polymer be matrix based on [Eu (fod) 3Dpbt] the preparation of fluorescent nano particles
With Eu (fod) 3Substitute Eu (tta) 3, press that method that document (Y.Wang, et al., Angew.Chem.Int.Ed.2004,43,5010) introduces is synthetic to obtain structural formula suc as formula the compound shown in the IV, with its called after [Eu (fod) 3Dpbt].
Figure G2009102034073D00111
(formula IV)
According to the method among the embodiment 1, with [the Eu (fod) of above-mentioned preparation 3Dpbt] alternative [Eu (tta) 3Dpbt] obtain solution A7, each solute concentration is in this solution: OTS, 2.0 * 10 -4Mol/L; PMMA, 5.0 * 10 -5Mol/L; [Eu (fod) 3Dpbt], 1.0 * 10 -4Mol/L; Substitute CTAB obtain solution B7 with dodecyl trimethoxy brometo de amonio (DTAB).According to preparation method among the embodiment 1 and condition, prepare mixture with PMMA, organo-siloxane and organosilicon polymer (OTS hydrolysis and by silicon hydroxyl condensation formation) and be substrate material, based on [Eu (fod) 3Dpbt] the colloidal sol of fluorescent nano particles, be designated as colloidal sol C7.
According to the method for embodiment 1, with colloidal sol C7 alternate embodiment 1 described colloidal sol C1, make the colloidal sol that the surface has the fluorescent nano particles of the amino active group that can supply graft reaction, be designated as colloidal sol E7.
The TEM test result shows that the distribution of sizes of fluorescent nano particles is 30～80nm among the colloidal sol E2 of above-mentioned preparation, and median size is 65nm.With prepared fluorescent nano particles centrifugation, with water washing, drying, with the OTS in its medium of acetone extract, gas-chromatography and icp analysis result show, the mol ratio of organo-siloxane OTS and Eu is 0.3 in the nanoparticle, hydrophobic polymer PMMA and Eu ionic mol ratio are 0.5 in the nanoparticle, and the mass percent content of organosilicon polymer is 15%.
The fluorescence spectrum test result shows that fluorescent nano particles has excellent visible light stimulated luminescence performance among the colloidal sol E7 of above-mentioned preparation, and its excitation peak in the visible region is positioned at 402nm, and afterbody is extended to 450nm.At wavelength is that the near-infrared laser of 832 nanometers excites down, and above-mentioned fluorescent nano particles can send bright ruddiness.
Embodiment 8: with the mixture of PMMA, organo-siloxane and organosilicon polymer be matrix based on [Eu (fod) 3Dpbt] the preparation of fluorescent nano particles
According to the method among the embodiment 1, with [Eu (fod) 3Dpbt] alternative [Eu (tta) 3Dpbt] obtain solution A8, each solute concentration is in this solution: OTS, 5.0 * 10 -5Mol/L; PMMA, 2.0 * 10 -4Mol/L; [Eu (fod) 3Dpbt], 1.0 * 10 -4Mol/L; Substitute CTAB obtain solution B8 with water-soluble polymer PVP, the concentration of PVP is 1 * 10 -4Mol/L.According to preparation method among the embodiment 1 and condition, prepare mixture with PMMA, organo-siloxane and organosilicon polymer (OTS hydrolysis and by silicon hydroxyl condensation formation) and be substrate material, based on [Eu (fod) 3Dpbt] the colloidal sol of fluorescent nano particles, be designated as colloidal sol C8.
The TEM test result shows that the distribution of sizes of fluorescent nano particles is 50～120nm among the colloidal sol C8 of above-mentioned preparation, and median size is 80nm.
With prepared fluorescent nano particles centrifugation, with water washing, drying, with the OTS in its medium of acetone extract, gas-chromatography and icp analysis result show, the mol ratio of OTS and Eu is 0.5 in the nanoparticle, hydrophobic polymer PMMA and Eu ionic mol ratio are 2 in the nanoparticle, and the quality percentage composition of organosilicon polymer is 4%.
The fluorescence spectrum test result shows that fluorescent nano particles has excellent visible light stimulated luminescence performance among the colloidal sol E7 of above-mentioned preparation, and its excitation peak in the visible region is positioned at 402nm, and afterbody is extended to 450nm.
Embodiment 9: with the mixture of polyethyl acrylate, organo-siloxane and organosilicon polymer be matrix based on [Eu (fod) 3Dmpbt] the preparation of fluorescent nano particles
The method of embodiment 7 that according to publication number is the patent of CN101066975A prepares structural formula suc as formula the title complex shown in the V, with its called after [Eu (fod) 3Dmpbt].
Figure G2009102034073D00131
(formula V)
With the tetrahydrofuran (THF) is solvent obtain solution A9, makes that each solute concentration is in the solution A 9: 3-(methacryloyl) propyl trimethoxy silicane, 1.0 * 10 -5Mol/L; Polyethyl acrylate, 1.0 * 10 -5Mol/L; [Eu (fod) 3Dmpbt], 1.0 * 10 -6Mol/L.Substitute CTAB with water-soluble polymer PVA, according to the method obtain solution B1 among the embodiment 1.Under room temperature and stirring condition; solution A 9 is added in the solution B 1; making the solution A 9 and the volume ratio of solution B 1 is 1: 5; prepare mixture with polyethyl acrylate, organo-siloxane and organosilicon polymer (hydrolysis of 3-(methacryloyl) propyl trimethoxy silicane and by silicon hydroxyl condensation formation) and be matrix, based on [Eu (fod) 3Dmpbt] fluorescent nano particles colloidal sol, be designated as colloidal sol C9.
With 0.1mL concentration is 2.5 * 10 -4The methanol solution D9 of the 3-of mol/L (2-aminoethyl amino) propyl trimethoxy silicane adds among the 10mL colloidal sol C9, in 40 ℃ of reactions 2 hours, centrifugation, the gained precipitation is scattered in the pure water again, makes the colloidal sol (being designated as colloidal sol E9) that the surface is connected to the fluorescent nano particles of the amino group that can supply graft reaction.
The TEM test result shows that the distribution of sizes of fluorescent nano particles is 20～50nm among the colloidal sol E7 of above-mentioned preparation, and median size is 35nm.With prepared fluorescent nano particles centrifugation, with water washing, drying; with the OTS in its medium of acetone extract; gas-chromatography and icp analysis result show; the mol ratio of organo-siloxane 3-(methacryloyl) propyl trimethoxy silicane and Eu is 0.05 in the nanoparticle; hydrophobic polymer P (St-co-MMA) is 0.1 with Eu ionic mol ratio in the nanoparticle, and the quality percentage composition of organosilicon polymer (hydrolysis of 3-(methacryloyl) propyl trimethoxy silicane and the network polymer that forms by the condensation of silicon hydroxyl) is 3%.
The fluorescence spectrum test result shows that fluorescent nano particles has excellent visible light stimulated luminescence performance among the colloidal sol E9 of above-mentioned preparation, and its excitation peak in the visible region is positioned at 409nm, and afterbody is extended to 460nm.Propanol solution with DCM is reference substance (Φ=0.57), is under the excitation light irradiation of 410nm at wavelength, and the rare earth ion fluorescence quantum yield that records fluorescent nano particles among the colloidal sol E6 of above-mentioned preparation is about 0.45.At wavelength is that the near-infrared laser of 800 nanometers excites down, and fluorescent nano particles can send bright ruddiness among the colloidal sol E9 of above-mentioned preparation, is that reference substance records its two-photon excitation action section and is about 50GM (1GM=10 with RhodamineB -50Cm 4S photo -1Molecule -1).
Embodiment 10: be the preparation based on the fluorescent nano particles of structure such as formula III europium complex of matrix with the mixture of polystyrene, organo-siloxane and organosilicon polymer
Being that the method for embodiment 6 of patent of CN101066975A is synthetic according to publication number obtains structural formula suc as formula the title complex shown in the VI.
With acetone is solvent, and obtain solution A10 makes that each solute concentration is in the solution A 10: dodecyl Trimethoxy silane (DTS), 5.0 * 10 -4Mol/L; Polystyrene, 5.0 * 10 -4Mol/L; Structure is suc as formula VI europium complex, 1.0 * 10 -3Mol/L; Compound concentration is 1.0 * 10 -3The aqueous solution B10 of the amphoteric ionic surfactant octadecyl trimethylammonium trimethyl-glycine of mol/L.Under room temperature and agitation condition, solution A 10 is added in the solution B 10, making the solution A 10 and the volume ratio of solution B 10 is 1: 5, prepare mixture with polystyrene, organo-siloxane and organosilicon polymer (DTS hydrolysis and by silicon hydroxyl condensation formation) and be matrix, fluorescent nano particles colloidal sol based on title complex VI is designated as colloidal sol C10.
Figure G2009102034073D00141
(formula VI)
With 0.1mL concentration is 1.0 * 10 -4The APS methanol solution D10 of mol/L adds among the 10mL colloidal sol C10, in 60 ℃ of reactions 1 hour, centrifugation is scattered in the gained precipitation in the pure water again, makes the colloidal sol (being designated as colloidal sol E10) that the surface is connected to the fluorescent nano particles of the amino group that can supply graft reaction.
The TEM test result shows that the distribution of sizes of the fluorescent nano particles among the colloidal sol E8 of above-mentioned preparation is 60～100nm, and median size is 85nm.With prepared fluorescent nano particles centrifugation, with water washing, drying, with the DTS in its medium of acetone extract, gas-chromatography and icp analysis result show that organo-siloxane DTS and Eu ionic mol ratio are 0.005 in the nanoparticle, and the mol ratio of PST and Eu is 0.5 in the nanoparticle.The quality percentage composition of organosilicon polymer (DTS hydrolysis and the network polymer that forms by the condensation of silicon hydroxyl) is 0.5%.
Nanoparticle among the colloidal sol E8 of above-mentioned preparation excites to descend can send bright ruddiness at visible light and near-infrared laser.
Embodiment 11: with the mixture of polymaleic acid dibutyl ester, organo-siloxane and organosilicon polymer be matrix based on [Eu (fod) 3Bpt] the preparation of fluorescent nano particles
The method of embodiment 7 that according to publication number is the patent of CN101066975A prepares structural formula suc as formula the title complex shown in the VII, with its called after [Eu (fod) 3Bpt].
With methyl alcohol is solvent obtain solution A11, makes that each solute concentration is in the solution A 11: 1H, 1H, 2H, 2H-fluoro octyl group Trimethoxy silane, 1.0 * 10 -2Mol/L; Polymaleic acid dibutyl ester, 1.0 * 10 -2Mol/L; [Eu (fod) 3Bpt], 1.0 * 10 -3Mol/L; Compound concentration is 1.0 * 10 -3The aqueous solution B11 of the anionic surfactant sodium dodecylbenzene sulfonate of mol/L.Under 0 ℃ and agitation condition, A11 solution is added in the B11 solution, making both volume ratios is 1: 9, prepare with polymaleic acid dibutyl ester, organo-siloxane and organosilicon polymer (1H, 1H, 2H, the hydrolysis of 2H-fluoro octyl group Trimethoxy silane and by silicon hydroxyl condensation formation) mixture be matrix, based on [Eu (fod) 3Bpt] the colloidal sol of fluorescent nano particles, be designated as colloidal sol C11.
With 0.1mL concentration is 1.0 * 10 -6The methanol solution D11 of the mercaptopropyl trimethoxysilane of mol/L adds among the 10mL colloidal sol C11, in 20 ℃ of reactions 12 hours, centrifugation is scattered in the gained precipitation in the pure water again, makes the colloidal sol (being designated as colloidal sol E11) that the surface is connected to the fluorescent nano particles of the mercapto groups that can supply graft reaction.
TEM result shows that the median size of the fluorescent nano particles among the colloidal sol C11 of above-mentioned preparation is 180nm.The distribution of sizes of the fluorescent nano particles among the colloidal sol E11 of above-mentioned preparation is 130～200nm, and median size is 180nm.Because the total mass of fluorescent nano particles among the add-on little colloidal sol C11 far away of APS, thus Electronic Speculum to characterize the size of nanoparticle in C11 colloidal sol that experiment records and the E11 colloidal sol very approaching.With prepared fluorescent nano particles centrifugation, with water washing, drying, with the 1H in its medium of acetone extract, 1H, 2H, 2H-fluoro octyl group Trimethoxy silane, gas-chromatography and icp analysis result show, organo-siloxane 1H in the nanoparticle, 1H, 2H, 2H-fluoro octyl group Trimethoxy silane and Eu ionic mol ratio are 2, and the mol ratio of polymaleic acid dibutyl ester and Eu is 0.1 in the nanoparticle.The quality percentage composition of organosilicon polymer (1H, 1H, 2H, hydrolysis of 2H-fluoro octyl group Trimethoxy silane and the network polymer that forms by the condensation of silicon hydroxyl) is 40%.
The fluorescence spectrum test result shows that the colloidal sol C11 and the fluorescent nano particles among the E11 of above-mentioned preparation all have good excited by visible light luminescent properties, and its excitation peak in the visible region is positioned at 410nm, and afterbody is extended to 460nm.At wavelength is that the near-infrared laser of 800nm excites down, and above-mentioned fluorescent nano particles can send bright ruddiness.
Figure G2009102034073D00151
(formula VII)
Embodiment 12: with the mixture of methylacrylic acid methyl esters styrol copolymer, organo-siloxane and organosilicon polymer be matrix based on [Eu (tta) 3Bpt] the preparation of fluorescent nano particles
Get N; N-diethyl para-bromoaniline (4mmol) is dissolved in the dry tetrahydrofuran (THF) of crossing logical argon gas deoxygenation; place-78 ℃ dry ice acetone bath; under argon shield, add 4mL n-BuLi (1.6M), slowly be warming up to room temperature after; continue to stir 30min; its adding is frozen in the three polychlorostyrene piperazines (5.4mmol) under-78 ℃, slowly be warming up to room temperature after, continue to stir 60min.Except that after desolvating, product separates with silica gel column chromatography, and eluent is a methylene dichloride.With the thick product of gained recrystallization in sherwood oil, obtain 2-(N, N-Diethyl Aniline-4-yl)-4, the yellow needle-like crystal 0.60g of 6-two chloro-1,3,5-triazines.Under argon shield, the potassium metal of newly cutting (1.25mmol) is added in the anhydrous tetrahydro furan; Add pyrazoles (1.75mmol), reflux 3h generates the pyrazoles negative ion; Reaction mixture is placed ice bath, to wherein adding 2-(N, N-Diethyl Aniline-4-yl)-4,6-two chloro-1,3,5-triazines (0.5mmol), stirring at room 1h, back flow reaction 8-9h in 80-85 ℃ oil bath then.Cooling, concentrate, with methylene dichloride and ethyl acetate mixed solution (volume ratio of methylene dichloride and ethyl acetate is 3: 1) make eluent, separate by silica gel column chromatography.The gained crude product is with mixed solution (volume ratio of sherwood oil and methylene dichloride is 1: the 1) recrystallization of sherwood oil and methylene dichloride, the yellow needle-like crystal 81mg of compound shown in the formula XIV.
Mass spectrum (EI MS) characterizes and records above-mentioned yellow needle-like crystal product molecular ion peak M/Z=360; Ultimate analysis value (quality percentage composition): C, 63.23% (63.32%); H, 5.58% (5.59%); N, 31.04% (31.09%), in the bracket theoretical value.X-ray analysis and nuclear magnetic resonance spectrum prove that above-mentioned yellow needle-like crystal product structure is suc as formula shown in the XIV.
Figure G2009102034073D00161
(formula XIV)
Under argon shield, with 10mL Eu (tta) 33H 2O (24.9 * 10 -3Mmol) tetrahydrofuran solution is added drop-wise to the compound (24.9 * 10 shown in the prepared formula XIV of 10mL -3Mmol) in the tetrahydrofuran solution, normal temperature stirs 30min down.Remove and desolvate,, separate out solid also washing, drying as precipitation agent, obtain the yellow powder 31mg of title complex shown in the formula VIII with normal hexane with a small amount of ether dissolution, filtration.
Figure G2009102034073D00162
(formula VIII)
Mass spectrum (MALDI-TOF MS) characterizes and records above-mentioned yellow powder product molecular ion peak M/Z=1176; Ultimate analysis (quality percentage composition): C, 43.80% (43.92%); H, 2.56% (2.74%); N, 9.41% (9.53%), in the bracket theoretical value; Prove that above-mentioned yellow powder product structure is suc as formula shown in the VIII.With [the Eu (tta) of title complex called after shown in the formula VIII 3Bpt].
With acetone is solvent obtain solution A12, makes that each solute concentration is in the solution A 12: phenyltrimethoxysila,e (PTS), 5.0 * 10 -6Mol/L; Styrene-methyl methacrylate copolymer, 1.0 * 10 -6Mol/L; [Eu (tta) 3Bpt], 1.0 * 10 -6Mol/L; Compound concentration is 1.0 * 10 -4Tween 20 aqueous solution B12 of mol/L.Under room temperature and agitation condition, solution A 12 is added in the solution B 12, making both volume ratios is 1: 4, make mixture with styrene-methyl methacrylate copolymer, organo-siloxane and organosilicon polymer (PTS hydrolysis and by silicon hydroxyl condensation formation) and be matrix, based on [Eu (tta) 3Bpt] the colloidal sol of fluorescent nano particles, be designated as colloidal sol C12.
With 0.1mL concentration is 1.0 * 10 -6The methanol solution D12 of the mercaptopropyl trimethoxysilane of mol/L adds among the 10mL colloidal sol C12, in 25 ℃ of reactions 12 hours, centrifugation was scattered in the gained precipitation in the pure water again, make the colloidal sol that the surface is connected to the fluorescent nano particles of the mercapto groups that can supply graft reaction, be designated as colloidal sol E12.
TEM result shows that the median size of the fluorescent nano particles among the colloidal sol C12 of above-mentioned preparation is 10nm.The distribution of sizes of the fluorescent nano particles among the colloidal sol E12 of above-mentioned preparation is 8～25nm, and median size is 10nm.Because the total mass of fluorescent nano particles among the add-on little colloidal sol C12 far away of APS, thus Electronic Speculum to characterize the size of nanoparticle in C10 colloidal sol that experiment records and the E10 colloidal sol very approaching.With prepared fluorescent nano particles centrifugation, with water washing, drying, with the PTS in its medium of acetone extract, gas-chromatography and icp analysis result show, the mol ratio of organo-siloxane PTS and Eu is 1.0 in the nanoparticle, hydrophobic polymer styrene-methyl methacrylate copolymer and Eu ionic mol ratio are 1.0 in the nanoparticle, and the mass percent content of organosilicon polymer is 20%.
The fluorescence spectrum test result shows that the colloidal sol C12 and the fluorescent nano particles among the E12 of above-mentioned preparation have good excited by visible light luminescent properties, and its excitation peak in the visible region is positioned at 422nm, and afterbody is extended to 475nm.
Embodiment 13: with the mixture of PS, PMMA, organo-siloxane and organosilicon polymer be matrix based on [Tb (dppc) 3] the preparation of fluorescent nano particles
Press document (H.
Figure G2009102034073D00171
Et al., Anal.Chem., 2005,77,2643) synthetic [Tb (dppc) that obtains of introduction method 3] (dppc=4-(2,4,6 ,-three last of the ten Heavenly stems oxygen base phenyl) pyridine-2, the 6-dicarboxylic acid).
With the tetrahydrofuran (THF) is solvent obtain solution A13, makes that each solute concentration is in the solution A 13: 1H, 1H, 2H, 2H-fluoro dodecyl triethoxysilicane, 1.0 * 10 -4Mol/L; PS, 2.0 * 10 -5Mol/L; PMMA, 2.0 * 10 -5Mol/L; [Tb (dppc) 3], 5.0 * 10 -5Mol/L.Solution A 1 with solution A 13 alternate embodiments 1, other conditions are with embodiment 1, make with PS, PMMA, organo-siloxane and organosilicon polymer (1H, 1H, 2H, the hydrolysis of 2H-fluoro dodecyl triethoxyl silane and by silicon hydroxyl condensation formation) mixture be matrix, based on [Tb (dppc) 3] the colloidal sol C13 of fluorescent nano particles.
With 0.1mL concentration is 1.0 * 10 -6The propyloic silanetriol of mol/L is received the methanol solution D13 of (carboxyethylsilanetriolsodium) and is added among the 10mL colloidal sol C13, in 0 ℃ of reaction 24 hours, centrifugation, again be scattered in the gained precipitation in the pure water, make the colloidal sol that the surface is connected to the fluorescent nano particles of the carboxylic group that can supply graft reaction, be designated as colloidal sol E13.
The TEM test result shows that the distribution of sizes of fluorescent nano particles is 60～120nm among the colloidal sol E13 of above-mentioned preparation, and median size is 90nm.The fluorescence spectrum test result shows that fluorescent nano particles has good ultraviolet excitation luminescent properties among the colloidal sol E13 of above-mentioned preparation, and its excitation peak is positioned at 320nm, and the emission main peak is positioned at 544nm.With prepared fluorescent nano particles centrifugation, with water washing, drying, with the 1H in its medium of acetone extract, 1H, 2H, 2H-fluoro dodecyl triethoxysilicane, gas-chromatography and icp analysis result show that the mol ratio of organo-siloxane and Eu is 0.2 in the nanoparticle, hydrophobic polymer PS, PMMA and Eu ionic mol ratio are 1.0 in the nanoparticle, and the mass percent content of organosilicon polymer is 10%.
Embodiment 14: with the mixture of PMMA, organo-siloxane and organosilicon polymer be matrix based on [Er (tta) 3Terpy] the preparation of fluorescent nano particles
Press the synthetic [Er (tta) that obtains of document (J.Luminescence 2007,127 for Y.H.Kim, et al., 707) introduction method 3Terpy] (terpy=terpyridyl).
With acetone is solvent obtain solution A14, and each solute concentration is in the solution A 14: OTS, 1.0 * 10 -2Mol/L; PMMA, 1.0 * 10 -2Mol/L; [Er (tta) 3Terpy], 1.0 * 10 -3Mol/L; Compound concentration is 1.0 * 10 -4The aqueous solution B14 of the nonionogenic tenside Triton X-100 of mol/L.Under 10 ℃ and agitation condition, solution A 14 is added in the solution B 14, make both volume ratios 1: 1, make mixture with PMMA, organo-siloxane and organosilicon polymer (OTS hydrolysis and by silicon hydroxyl condensation formation) and be matrix, based on [Er (tta) 3Terpy] the colloidal sol of fluorescent nano particles, be designated as colloidal sol C14.
According to the method for embodiment 1, with colloidal sol C14 alternate embodiment 1 described colloidal sol C1, make the colloidal sol that the surface has the fluorescent nano particles of the amino active group that can supply graft reaction, be designated as colloidal sol E14.
The TEM test result shows that the distribution of sizes of fluorescent nano particles is 50～100nm among the colloidal sol E14 of above-mentioned preparation, and median size is 70nm.The fluorescence spectrum test result shows, is under the optical excitation of 355nm at wavelength, and the fluorescent nano particles of above-mentioned preparation has good near-infrared luminous performance, and its near infrared emission main peak is positioned at 1530nm.
Embodiment 15: with the mixture of styrene-methyl methacrylate copolymer, organo-siloxane and organosilicon polymer be matrix based on [Yb (hfac) 3PBA] the preparation of fluorescent nano particles
Press document (J.A.Weinstein, et al., Dalton Trans., 2007,1484-1491; Y.Hasegawa, J.Phys.Chem.1996,100,10201) synthetic [Yb (hfac) that obtains of introduction method 3PBA] (hfac=1,1,1,5,5,5-hexafluoro-pentane-2,4-diketone negative ion, PBA=1-(9-methylene radical-anthracene)-2-(2-pyridine) benzoglyoxaline).
With acetone is solvent obtain solution A15, and each solute concentration is in the solution A 15: OTS, 1.0 * 10 -3Mol/L; Styrene-methyl methacrylate copolymer, 5.0 * 10 -4Mol/L; [Yb (hfac) 3PBA], 2.0 * 10 -4Mol/L.Solution A 1 with solution A 15 alternate embodiments 1, other conditions are with embodiment 1, make mixture with styrene-methyl methacrylate copolymer, organo-siloxane and organosilicon polymer (OTS hydrolysis and by silicon hydroxyl condensation formation) and be matrix, based on [Yb (hfac) 3PBA] the colloidal sol of fluorescent nano particles, be designated as colloidal sol C15.
According to the method for embodiment 1, with colloidal sol C15 alternate embodiment 1 described colloidal sol C1, make surperficial colloidal sol with described fluorescent nano particles of the amino active group that can supply graft reaction, be designated as colloidal sol E15.
TEM result shows that the distribution of sizes of the fluorescent nano particles among the colloidal sol E15 of above-mentioned preparation is 60～120nm, and median size is 80nm.With prepared fluorescent nano particles centrifugation, with water washing, drying, with the OTS in its medium of acetone extract, gas-chromatography and icp analysis result show, the mol ratio of organo-siloxane OTS and Yb is 0.5 in the nanoparticle, hydrophobic polymer styrene-methyl methacrylate copolymer and Yb ionic mol ratio are 0.1 in the nanoparticle, and the mass percent content of organosilicon polymer is 40%.This fluorescence spectrum test result shows, is under the optical excitation of 380nm at wavelength, and the fluorescent nano particles among the colloidal sol E15 of above-mentioned preparation has good near infrared light luminosity, and its near infrared emission main peak is positioned at 980nm.
Embodiment 16: with the mixture of PMMA, organo-siloxane and organosilicon polymer be matrix based on [Nd (tta) 3BPTZ] the preparation of fluorescent nano particles
Press document (M.Ward, et al., Dalton Trans., 2003, the 808-814) synthetic [Nd (tta) that obtains of introduction method 3BPTZ] (BPTZ=3, two (2-pyridyl) tetrazines of 6-).
With acetone is solvent obtain solution A16, and each solute concentration is in the solution A 16: OTS, 1.0 * 10 -2Mol/L; PMMA, 1.0 * 10 -2Mol/L; [Nd (tta) 3BPTZ], 1.0 * 10 -3Mol/L; Compound concentration is 1.0 * 10 -4The water-soluble polymer PVA aqueous solution B16 of mol/L.Under 20 ℃ and agitation condition, solution A 16 is added in the solution B 16, making both volume ratios is 1: 50; Make mixture with PMMA, organo-siloxane and organosilicon polymer (OTS hydrolysis and by silicon hydroxyl condensation formation) and be matrix, based on [Nd (tta) 3BPTZ] the colloidal sol of fluorescent nano particles, be designated as colloidal sol C16.
According to the method for embodiment 1, with colloidal sol C16 alternate embodiment 1 described colloidal sol C1, making the surface can be the colloidal sol of the described fluorescent nano particles of amino for the active group of graft reaction, is designated as colloidal sol E16.
The TEM test result shows that the distribution of sizes of the fluorescent nano particles among the colloidal sol E16 of above-mentioned preparation is 40～80nm, and median size is 65nm.With prepared fluorescent nano particles centrifugation, with water washing, drying, with the OTS in its medium of acetone extract, gas-chromatography and icp analysis result show, the mol ratio of organo-siloxane OTS and Nd is 0.3 in the nanoparticle, hydrophobic polymer PMMA and Nd ionic mol ratio are 0.1 in the nanoparticle, and the mass percent content of organosilicon polymer is 28%.The fluorescence spectrum test result shows, is under the optical excitation of 337nm at wavelength, and the fluorescent nano particles in the colloidal sol of above-mentioned preparation has good near-infrared luminous performance near 1060nm.
Embodiment 17: with the mixture of styrene-methyl methacrylate copolymer, organo-siloxane and organosilicon polymer be matrix based on [Eu (tta) 3Dpbt] preparation of fluorescent nano particles
With OTS, APS, copolymer of methyl methacrylatestyrene and [Eu (tta) 3Dpbt] be dissolved in the acetone, being mixed with OTS concentration is 2.3 * 10 -3Mol/L, APS concentration is 4.6 * 10 -5Mol/L, styrene-methyl methacrylate copolymer concentration is 2.9 * 10 -6Mol/L, [Eu (tta) 3Dpbt] concentration is 2.3 * 10 -4The acetone soln of mol/L is designated as solution F17.Under room temperature and agitation condition, the concentration that 1.5mL solution F17 injection is added 7.5mL is 1.0 * 10 -3Among the nonionogenic tenside Triton X-100 (being designated as solution B 17) of mol/L (solution F17 and solution B 17 volume ratios are 1: 5), stir 0.5hr, the colloidal sol that reaction is obtained is crossed Sephadex G-25 chromatographic column and is removed acetone and unnecessary tensio-active agent (is elutriant with water), finally obtain being dispersed in the stable sol in the water, making promptly that the surface has can be for the amino active group of graft reaction, copolymer of methyl methacrylatestyrene, the mixture of organo-siloxane and organosilicon polymer (OTS and APS hydrolysis also form by the condensation of silicon hydroxyl) is a matrix, based on [Eu (tta) 3Dpbt] the colloidal sol of fluorescent nano particles, be designated as colloidal sol G17.
The TEM test result shows that the distribution of sizes of fluorescent nano particles is 10～50nm among the colloidal sol G17 of above-mentioned preparation, and median size is 27nm (among Fig. 1 shown in a).With prepared fluorescent nano particles centrifugation, with water washing, drying, with the OTS in its medium of acetone extract, gas-chromatography and icp analysis result show, the mol ratio of organo-siloxane OTS and Eu is 0.6 in the nanoparticle, hydrophobic polymer styrene-methyl methacrylate copolymer and Eu ionic mol ratio are 0.01 in the nanoparticle, and the mass percent content of organosilicon polymer is 25%.The fluorescence spectrum test result shows that the fluorescent nano particles among the colloidal sol G17 of above-mentioned preparation has good excited by visible light luminescent properties, and its excitation peak in the visible region is positioned at 412nm, and afterbody is extended to 470nm.
Embodiment 18: with the mixture of acrylic styrene copolymer, organo-siloxane and organosilicon polymer be matrix based on [Eu (tta) 3Dmpbt] preparation of fluorescent nano particles
Be that the embodiment 1 of patent of CN101070318A and the method for embodiment 2 prepare the [Eu (tta) of the compound shown in the formula IX according to publication number 3Dmpbt].
With acetone is solvent obtain solution F18, and each solute concentration is among the solution F18: DTS, 1.0 * 10 -6Mol/L; APS concentration is 4.6 * 10 -5Mol/L; Methyl acrylate-styrol copolymer, 1.0 * 10 -6Mol/L; [Eu (tta) 3Dmpbt], 1.0 * 10 -6Mol/L; Compound concentration is 1.0 * 10 -3Tween 20 aqueous solution B18 of mol/L.Solution F18 is added among the B18, and making both volume ratios is 1: 3; Under 0 ℃ and agitation condition, react 24hr, making that the surface has can be for the amino active group of graft reaction, with the mixture of methyl acrylate styrol copolymer, organo-siloxane and organosilicon polymer (OTS hydrolysis and by silicon hydroxyl condensation formation) is matrix, based on [Eu (tta) 3Dmpbt] fluorescent nano particles colloidal sol G18.
The TEM test result shows that the fluorescent nano particles distribution of sizes among the colloidal sol G18 of above-mentioned preparation is 20～50nm, and median size is 31nm.With prepared fluorescent nano particles centrifugation, with water washing, drying, with the DTS in its medium of acetone extract, gas-chromatography and icp analysis result show, the mol ratio of organo-siloxane DTS and Eu is 0.5 in the nanoparticle, hydrophobic high molecular weight acrylic ester-styrol copolymer and Eu ionic mol ratio are 1 in the nanoparticle, and the mass percent content of organosilicon polymer is 40%.The fluorescence spectrum test result shows that the fluorescent nano particles among the colloidal sol G18 of above-mentioned preparation has good excited by visible light luminescent properties, and its excitation peak in the visible region is positioned at 409nm, and afterbody is extended to 458nm.
Figure G2009102034073D00211
(formula IX)
Embodiment 19: with the mixture of toxilic acid dibutyl ester-styrol copolymer, organo-siloxane and organosilicon polymer be matrix based on the preparation of structure suc as formula the fluorescent nano particles of IV title complex
The method of embodiment 10 that according to publication number is the patent of CN101066975A prepares structural formula suc as formula the title complex shown in the X.
With the tetrahydrofuran (THF) is solvent preparation F19, and each solute concentration is among the solution F19: PTS, 1.0 * 10 -3Mol/L; APS, 1.0 * 10 -4Mol/L; Toxilic acid dibutyl ester-styrol copolymer, 1.0 * 10 -4Mol/L; Structure is suc as formula the europium complex of X, 2.0 * 10 -4Mol/L; Compound concentration is 1.0 * 10 -3 Tween 40 aqueous solution B19 of mol/L.Solution F19 is added among the B19, and making both volume ratios is 1: 50; Under 80 ℃ and agitation condition, react 0.5hr, making the surface can be amino for the active group of graft reaction, the mixture of toxilic acid dibutyl ester-styrol copolymer, organo-siloxane and organosilicon polymer (PTS and APS hydrolysis also form by the condensation of silicon hydroxyl) is a matrix, based on the fluorescent nano particles colloidal sol of structure, be designated as colloidal sol G19 suc as formula the title complex of IV.
TEM result shows that the distribution of sizes of the fluorescent nano particles among the colloidal sol G19 of above-mentioned preparation is 60～110nm, and median size is 85nm.With prepared fluorescent nano particles centrifugation, with water washing, drying, with the PTS in its medium of acetone extract, gas-chromatography and icp analysis result show, the mol ratio of organo-siloxane PTS and Eu is 0.2 in the nanoparticle, hydrophobic polymer toxilic acid dibutyl ester-styrol copolymer and Eu ionic mol ratio are 0.5 in the nanoparticle, and the mass percent content of organosilicon polymer is 15%.Nanoparticle among the colloidal sol G19 of above-mentioned preparation can send ruddiness under visible light and near-infrared laser excite.
Figure G2009102034073D00212
(formula X)
Embodiment 20: with the mixture of PMMA, organo-siloxane, organo-siloxane and organosilicon polymer be matrix based on [Eu (fod) 3Dpbt] preparation of complex fluorescent nanoparticle
With acetone is solvent obtain solution F20, and each solute concentration is among the solution F20: isobutyl-Trimethoxy silane, 1.0 * 10 -5Mol/L; 3-(2-aminoethyl amino) propyl trimethoxy silicane, 1.0 * 10 -6Mol/L; PMMA, 1.0 * 10 -5Mol/L; [Eu (fod) 3Dpbt], 2.0 * 10 -4Mol/L; Compound concentration is 1.0 * 10 -3The amphoteric ionic surfactant sodium dodecyl aminopropionitrile aqueous solution B20 of mol/L.Solution F20 is added among the B20, and making both volume ratios is 1: 1; Under 40 ℃ and agitation condition, react 6hr, make to such an extent that surperficial active group for graft reaction is amino, the mixture of PMMA, organo-siloxane and organosilicon polymer (isobutyl-Trimethoxy silane and 3-(2-aminoethyl amino) propyl trimethoxy silicane hydrolysis also forms by the condensation of silicon hydroxyl) is a matrix, based on [Eu (fod) 3Dpbt] fluorescent nano particles colloidal sol, be designated as colloidal sol G20.
TEM result shows that the distribution of sizes of fluorescent nano particles is 40～90nm among the colloidal sol G20 of above-mentioned preparation, and median size is 65nm.With prepared fluorescent nano particles centrifugation, with water washing, drying, with the isobutyl-Trimethoxy silane in its medium of acetone extract, gas-chromatography and icp analysis result show, the mol ratio of organo-siloxane isobutyl-Trimethoxy silane and Eu is 0.1 in the nanoparticle, hydrophobic polymer PMMA and Eu ionic mol ratio are 0.05 in the nanoparticle, and the mass percent content of organosilicon polymer is 8%.The fluorescence spectrum test result shows that the fluorescent nano particles among the colloidal sol G20 of above-mentioned preparation has good excited by visible light luminescent properties, and its excitation peak in the visible region is positioned at 402nm, and afterbody is extended to 450nm.
Embodiment 21: be shell with silicon-dioxide, to be surrounded by [Eu (tta) 3Dpbt] the particle of PMMA-organosilicon polymer mixture be the preparation of the shell-core structure nanoparticle of kernel
Preparing PMMA, organo-siloxane and organosilicon polymer according to the method for embodiment 1 is matrix, based on [Eu (tta) 3Dpbt] the colloidal sol C1 of nanoparticle.
With 0.2mL concentration is among the above-mentioned colloidal sol C1 of tetramethoxy-silicane (TMOS) methanol solution H14 adding 10mL of 0.5mol/L, 60 ℃ were reacted 2 hours, and centrifugation is scattered in the gained precipitation in the 10mL water, making shell is silicon dioxide layer, and kernel is for being surrounded by title complex [Eu (tta) 3Dpbt] the colloidal sol of fluorescent nano particles of particle of mixture of PMMA, organo-siloxane and organosilicon polymer, be designated as colloidal sol J21.
With 0.1mL concentration is 3.7 * 10 -5The APS methanol solution D21 of mol/L joins among the colloidal sol J21 of the above-mentioned preparation of 10mL, in 60 ℃ of reactions 1 hour, centrifugation, again be scattered in the gained precipitation in the water, make the surface and have amino group, PMMA and organosilicon polymer are inner core matrix, based on [Eu (tta) 3Dpbt] the colloidal sol of core-shell structured fluorescence nanoparticle, be designated as colloidal sol K21.
The TEM test result shows that the median size of shell-core structure fluorescent nano particles is 84nm among the above-mentioned preparation colloidal sol J21.The distribution of sizes of shell-core structure fluorescent nano particles is 50～100nm among the colloidal sol K21 of above-mentioned preparation, and median size is 85nm.Electronic Speculum and EDX analysis revealed, prepared nanoparticle have the silicon oxide shell that thickness is 5 nanometers.With prepared fluorescent nano particles centrifugation, with water washing, drying, with the OTS in its medium of acetone extract, gas-chromatography and icp analysis result show, organo-siloxane OTS and Eu mol ratio are 0.4 in the nanoparticle, hydrophobic polymer PMMA and Eu ionic mol ratio are 0.1 in the fluorescent nano particles, and the quality percentage composition of organosilicon polymer is 40%.
The fluorescence spectrum test result shows that fluorescent nano particles has good excited by visible light luminescent properties among the colloidal sol J21 of above-mentioned preparation and the K21, and its excitation peak in the visible region is positioned at 412nm, and afterbody is extended to 480nm.At wavelength is that the near-infrared laser of 832 nanometers excites down, and this fluorescent nano particles can send bright ruddiness.
Embodiment 22: be shell with silicon-dioxide, to be surrounded by [Eu (tta) 3Dpbt] the particle of PMMA-organosilicon polymer mixture be the preparation of the shell-core structure nanoparticle of kernel
Is matrix according to the method for embodiment 8 preparation with PMMA, organo-siloxane and organosilicon polymer, based on [Eu (fod) 3Dpbt] the colloidal sol C8 of nanoparticle.
With 1.0mL concentration is 5.0 * 10 -1The tetraethoxysilane of mol/L (TEOS) ethanolic soln H20 adds among the above-mentioned colloidal sol C8 of 10mL, 80 ℃ of reactions 1 hour, and centrifugation will precipitate and be scattered in again in the 10mL water, and making shell is silicon dioxide layer, and kernel is for being surrounded by title complex [Eu (fod) 3Dpbt] the colloidal sol of fluorescent nano particles of particle of mixture of PMMA, organo-siloxane and organosilicon polymer, be designated as colloidal sol J22.
With 0.1mL concentration is 1.0 * 10 -5The sulfydryl propyl trimethoxy silicane methanol solution (being designated as solution D 22) of mol/L adds among the colloidal sol J22 of the above-mentioned preparation of 10mL, 20 ℃ were reacted 24 hours, centrifugation, precipitation is scattered in the water, make the surface and have mercapto groups, with silicon-dioxide is shell, and PMMA and organosilicon polymer are inner core matrix, based on [Eu (fod) 3Dpbt] the colloidal sol of core-shell structured fluorescence nanoparticle, be designated as colloidal sol K22.
The TEM test result shows that the distribution of sizes of the shell-core structure fluorescent nano particles among the colloidal sol K22 of above-mentioned preparation is 50～90nm, and median size is 60nm.Electronic Speculum and EDX analysis revealed, the nanoparticle among the colloidal sol K22 of above-mentioned preparation has the silicon oxide shell that mean thickness is about 20 nanometers.The fluorescence spectrum test result shows that the fluorescent nano particles among the colloidal sol K22 of above-mentioned preparation has good excited by visible light luminescent properties, and its excitation peak in the visible region is positioned at 402nm, and afterbody is extended to 450nm.
Embodiment 23: be shell with silicon-dioxide, to be surrounded by [Eu (fod) 3Dmpbt] the particle of acrylate styrol copolymer-organosilicon polymer mixture be the preparation of the shell-core structure nanoparticle of kernel
Is matrix according to the method for embodiment 9 preparation with polyethyl acrylate, organo-siloxane and organosilicon polymer, based on [Eu (fod) 3Dmpbt] the colloidal sol C9 of nanoparticle.
With 1.0mL concentration is 5.0 * 10 -1The TEOS ethanolic soln H20 of mol/L adds among the above-mentioned 10mL colloidal sol C9, and 40 ℃ were reacted 6 hours, and centrifugation is scattered in the gained precipitation in the 10mL water, and making shell is silicon dioxide layer, and kernel is for being surrounded by title complex [Eu (fod) 3Dpbt] the colloidal sol of fluorescent nano particles of particle of mixture of polyethyl acrylate, organo-siloxane and organosilicon polymer, be designated as colloidal sol J23.
With 0.1mL concentration is 1.0 * 10 -4The 3-of mol/L (2-aminoethyl amino) propyl trimethoxy silicane methanol solution D23 adds among the 10mL colloidal sol J23,40 ℃ were reacted 6 hours, centrifugation, the gained precipitation is scattered in the water, make the surface and have amino group, with silicon-dioxide is shell, and polyethyl acrylate-organosilicon polymer mixture is an inner core matrix, based on [Eu (fod) 3Dpbt] the colloidal sol of core-shell structured fluorescence nanoparticle, be designated as colloidal sol K23.
The TEM test result shows that the distribution of sizes of the shell-core structure fluorescent nano particles among the colloidal sol K23 of above-mentioned preparation is 35～65nm, and median size is 50nm.With prepared fluorescent nano particles centrifugation, with water washing, drying, with the OTS in its medium of acetone extract, gas-chromatography and icp analysis result show, the mol ratio of OTS and Eu is 0.3 in the nanoparticle, hydrophobic polymer PMMA and Eu ionic mol ratio are 2 in the nanoparticle, and the quality percentage composition of organosilicon polymer is 10%.Electronic Speculum and EDX analysis revealed, the nanoparticle among the colloidal sol K21 of above-mentioned preparation has the silicon oxide shell that mean thickness is about 7 nanometers.The fluorescence spectrum test result shows that the fluorescent nano particles among the colloidal sol K23 of above-mentioned preparation has good excited by visible light luminescent properties, and its excitation peak in the visible region is positioned at 409nm, and afterbody is extended to 460nm.
Embodiment 24: be shell with silicon-dioxide, to be surrounded by [Eu (tta) 3Bpt] the particle of styrene-methyl methacrylate copolymer-organosilicon polymer mixture be the preparation of the shell-core structure nanoparticle of kernel
The mixture for preparing styrene-methyl methacrylate copolymer, organo-siloxane and organosilicon polymer according to the method for embodiment 12 is a matrix, based on [Eu (tta) 3Bpt] the colloidal sol C12 of fluorescent nano particles.
With 0.1mL concentration is 5.0 * 10 -2The TMOS ethanolic soln H24 of mol/L adds among the above-mentioned colloidal sol C12 of 10mL, and 0 ℃ was reacted 24 hours down, and centrifugation is scattered in the gained precipitation in the 10mL pure water again, and making shell is silicon dioxide layer, and kernel is for being surrounded by title complex [Eu (tta) 3Bpt] the colloidal sol of fluorescent nano particles of particle of mixture of styrene-methyl methacrylate copolymer, organo-siloxane and organosilicon polymer, be designated as colloidal sol J24.
With 0.1mL concentration is 1.0 * 10 -4The aqueous solution D24 that the propyloic silanetriol of mol/L is received adds among the 10mL colloidal sol J24,0 ℃ was reacted 24 hours, centrifugation, the gained precipitation is scattered in the water, make the surface and have carboxylic group, with silicon-dioxide is shell, and styrene-methyl methacrylate copolymer-organosilicon polymer is an inner core matrix, based on [Eu (tta) 3Bpt] the colloidal sol of core-shell structured fluorescence nanoparticle be designated as colloidal sol K24.
The TEM test result shows that the fluorescent nano particles median size is 12nm among the colloidal sol J24 of above-mentioned preparation.The fluorescent nano particles distribution of sizes is 10～27nm among the colloidal sol K24 of above-mentioned preparation, and median size is 12nm.Because the total mass of fluorescent nano particles among the add-on little colloidal sol J24 far away that receives of propyloic silanetriol, thus Electronic Speculum to characterize the size of nanoparticle in J24 colloidal sol that experiment records and the K24 colloidal sol very approaching.Electronic Speculum and EDS analysis revealed, the nanoparticle among the colloidal sol K24 of above-mentioned preparation has the silicon oxide shell that mean thickness is about 1 nanometer.With prepared fluorescent nano particles centrifugation, with water washing, drying, with the PTS in its medium of acetone extract, gas-chromatography and icp analysis result show, the mol ratio of organo-siloxane PTS and Eu is 0.5 in the nanoparticle, hydrophobic polymer styrene-methyl methacrylate copolymer and Eu ionic mol ratio are 1.0 in the nanoparticle, and the mass percent content of organosilicon polymer is 25%.
The fluorescence spectrum test shows, the colloidal sol J24 and the fluorescent nano particles among the K24 of above-mentioned preparation have excellent visible light stimulated luminescence performance, and its excitation peak in the visible region is positioned at 422nm, and afterbody is extended to 475nm.
Embodiment 25: be shell with silicon-dioxide, to be surrounded by [Eu (fod) 3Bpt] the particle of polymaleic acid dibutyl ester-organosilicon polymer mixture be the preparation of the shell-core structure nanoparticle of kernel
Is matrix according to the preparation of the method for embodiment 11 with the mixture of polymaleic acid dibutyl ester, organo-siloxane and organosilicon polymer, based on [Eu (fod) 3Bpt] fluorescent nano particles colloidal sol C11.
With 1.0mL concentration is 5.0 * 10 -1The TMOS ethanolic soln H25 of mol/L adds among the above-mentioned colloidal sol C11 of 10mL, 80 ℃ were reacted 1 hour, centrifugation, the gained precipitation is scattered in the 10mL pure water again, make the surface and have oh group, with silicon-dioxide is shell, and polymaleic acid dibutyl ester-organosilicon polymer is an inner core matrix, based on [Eu (fod) 3Bpt] the colloidal sol of core-shell structured fluorescence nanoparticle, be designated as colloidal sol J25.
The TEM test result shows that the fluorescent nano particles distribution of sizes is 150～220nm among the colloidal sol J25 of above-mentioned preparation, and median size is 200nm.Electronic Speculum and EDX analysis revealed, the nanoparticle among the colloidal sol J25 of above-mentioned preparation has the silicon oxide shell that mean thickness is about 10 nanometers.With prepared fluorescent nano particles centrifugation, with water washing, drying, with the 1H in its medium of acetone extract, 1H, 2H, 2H-fluoro octyl group Trimethoxy silane, gas-chromatography and icp analysis result show, organo-siloxane 1H in the nanoparticle, 1H, 2H, 2H-fluoro octyl group Trimethoxy silane and Eu ionic mol ratio are 1, and the mol ratio of polymaleic acid dibutyl ester and Eu is 0.1 in the nanoparticle.The quality percentage composition of organosilicon polymer (1H, 1H, 2H, hydrolysis of 2H-fluoro octyl group Trimethoxy silane and the network polymer that forms by the condensation of silicon hydroxyl) is 40%.Fluorometric investigation shows that the fluorescent nano particles among the colloidal sol J25 of above-mentioned preparation has good excited by visible light luminescent properties, can send bright ruddiness under excited by visible light.
Embodiment 26: the synthetic and compliance test result of nano fluorescent bioprobe (the anti-human IgG of fluorescent nanometer particle to mark goat)
With embodiment 1 prepared fluorescent nano particles is the anti-human IgG of label particles mark goat.Concrete grammar is as described below:
With the 0.1mL mass percentage concentration is that 1% glutaraldehyde water solution adds among the fluorescent nano particles colloidal sol E1 of 2mL embodiment 1 preparation, stirring at room 1hr, centrifugation, to precipitate with Tris buffered soln (pH=7.5,50mM) be scattered in after the washed twice in the Tris buffered soln of 2mL, make colloidal sol through glutaraldehyde activatory fluorescent nano particles.In this colloidal sol, add 20 μ L concentration and be 70～80 μ g/mL (be designated as IgG-FITC, room temperature reaction 2hr adds 2mg bovine serum albumin (BSA) again and continues reaction 2h, adds 1mg NaBH then by the anti-human IgG of FITC labelled goat 4Reaction 1hr.With the reaction solution centrifugation that obtains, be scattered in after precipitation is washed 2 times with Tris buffered soln and contain bovine serum albumin (the quality percentage composition is 0.1%) and NaN 3In the Tris buffered soln of (the quality percentage composition is 0.01%), promptly obtain nano fluorescent bioprobe (the anti-human IgG of fluorescent nanometer particle to mark goat) colloid.
The fluorescence spectrum test result shows, the fluorescence that is connected to the FITC among the IgG-FITC of nanoparticle surface can be detected its excitation spectrum (λ Em=530nm) and emission spectrum (λ Ex=470nm) as shown in Figure 4.This experimental result shows that the anti-human IgG of goat has been covalently bonded to described fluorescent nano particles surface, has constituted the fluoroimmunoassay nano-probe.FITC and [Eu (tta) in the nano-probe 3Dpbt] excitation spectrum and emission spectrum as shown in Figure 4, Eu ion fluorescence emission peak is sharp-pointed and have a very big Stokes shift, and the FITC fluorescence emission peak is very wide and excitation spectrum and emission spectrum are overlapping bigger, Eu ion fluorescence peak height is about 80 times of FITC fluorescence peak height, explanation is on luminescent properties, and fluorescent nano particles probe of the present invention is much better than traditional organic dye probe.Among Fig. 4, left side figure be fluorescent nano particles with the nanoparticle probe of acquisition after the anti-human IgG of organic fluorescent dye FITC labelled goat combines in [Eu (tta) 3Dpbt] excitation spectrum (A among the figure) (λ Em=614nm) with emission spectrum (B among the figure) (λ Ex=412nm); Right figure is excitation spectrum (A among the figure) (λ of FITC Em=530nm) with emission spectrum (B among the figure) (λ Ex=470nm).
Embodiment 27: nano fluorescent bioprobe (the anti-human IgG of fluorescent nanometer particle to mark goat) synthetic
With embodiment 2 prepared fluorescent nano particles is the anti-human IgG of marker mark goat.Concrete grammar is as described below:
With among the fluorescent nano particles colloidal sol C1 of embodiment 2 preparation 40, centrifugal under the 000g, the gained precipitation is scattered in the water again.This process repeats twice to remove excessive Surfactant CTAB; Fluorescent nano particle is precipitated (pH=7.8,10mmol L in the Tris-HCl buffered soln that is scattered in 8mL -1), be that the anti-human IgG solution of goat of 10mg/mL mixes with 0.1mL concentration then, and shook 2 hours in 25 ℃ of constant temperature, make the anti-human IgG of goat be adsorbed in the mixture (being designated as anti-IgG ◎ LNP) that the fluorescent nano particles surface forms anti-human IgG of goat and fluorescent nano particles.Centrifugation, precipitation is scattered in the 8mL Tris-HCl buffered soln resulting anti-IgG ◎ LNP stand-by with Tris-HCl buffered soln washed twice.The colloidal solution of this bioprobe can send Eu ionic characteristic radiation under the optical excitation of 420nm, the strongest emission wavelength is 614nm.
Embodiment 28: the compliance test result of the immunodetection that is used for human IgG of the anti-human IgG of fluorescent nanometer particle to mark goat
Sheet glass is put into the vitriol oil to be soaked 1 day, put into saturated aqueous sodium hydroxide solution behind the deionized water rinsing and soaked 1 day, rinse well with deionized water once more, put into acetone and ethanol supersound process 15min successively, dry up with nitrogen behind the alcohol flushing, obtain the glass substrate of cleaning.It is that the APS methanol solution of 10mmol/L soaks 12h that this cleaning substrate is put into concentration, takes out the back and dries up with alcohol flushing, nitrogen, obtains the substrate of surface amino groups silanization.It is 5% glutaraldehyde water solution that the substrate of this surface amino groups silanization is put into mass percentage concentration, room temperature reaction 12h, take out back PBST buffered soln (50mM, pH=7.4, tween 200.05%) flushing repeatedly, rinse well with deionized water, nitrogen dries up, and makes the substrate of surperficial aldehyde radicalization.(50mM, pH=7.4) point sample is put into wet box and is hatched 2h for 37 ℃ on the substrate of the surperficial aldehyde radicalization of above-mentioned preparation will to contain the PBS buffered soln of 1.0mg/mL human IgG.Then, with PBS buffered soln (50mM, pH=7.4) wash this substrate three times after, put it into again contain the PBS buffered soln that mass percentage concentration is 1% BSA (50mM, pH=7.4) in, hatch 2h in 37 ℃.(50mM pH=7.4) after the rinse substrate three times, puts it in the colloidal solution of the prepared fluoroimmunoassay nano-probe of embodiment 26, hatches 1h in 37 ℃ with PBS buffered soln.Sample after the nano-probe identification is taken out the back wash three times with Tris-Tween20 buffered soln (contain the quality percentage composition and be 0.05% Tween20, pH is 7.5, the Tris solution of 50mM), wash repeatedly with deionized water, nitrogen dries up.
Above-mentioned reacted substrate is observed under fluorescent microscope, the result shows, at wavelength is under the optical excitation of 406nm, can see the bright ruddiness that sends on the position behind the human IgG point sample, shows that institute's synthetic fluoroimmunoassay nano-probe has the excellent specificity recognition function to human IgG.
Embodiment 29: adopt anti-IgG@LNP as fluorescent probe, detect human IgG with solid phase interlayer fluoroimmunoassay.
The anti-human IgG of goat is scattered in the carbonate buffer solution of pH=9.16, and making its concentration is 20 μ g mL -1The anti-human IgG solution of this goat is added in the enzyme plate, and every hole add-on is 100 μ L.Allow this enzyme plate 4 ℃ of incubated overnight.After pouring out the anti-human IgG solution of goat, with Tris-HCl-Tween buffered soln (pH=7.8,10mmolL -1, contain 0.5%Tween-20) and cleaning of enzyme target 3 times.Afterwards, the Tris-HCl-Tween buffered soln of BSA (1.0%) is added in the above-mentioned enzyme plate hole, every hole add-on is 150 μ L, and this enzyme plate is placed in the constant temperature mixing instrument in 25 ℃ of insulation 1.5h.Pour out BSA solution, use Tris-HCl-Tween buffered soln cleaning of enzyme target 3 times, adding concentration in different holes respectively is the human IgG solution of 0.1ng/mL to 10 μ g/mL, and in 37 ℃ of incubation 1.5h..Pour out human IgG solution, use Tris-HCl-Tween buffered soln cleaning of enzyme target 3 times.The anti-IgG@LNP of fluorescent probe that adds example 27 preparations, and in 37 ℃ of incubation 1.5h.At last excessive fluorescent probe solution is poured out, and with Tris-HCl-Tween buffered soln cleaning of enzyme target 3 times.
The light that with the wavelength is 415nm is exciting light, adopts multi-functional microplate reader to measure the fluorescence intensity in each hole of enzyme plate, and the result shows that when human IgG concentration was in 5ng/mL～5 μ g/mL scopes, measured fluorescence intensity was with linear increase (Fig. 5) of human IgG concentration.The detection of this fluorescence immunoassay fluorometric analysis is limited to 15ng/mL.

Claims (21)

1. fluorescent nano particles comprises substrate material and the fluorescence dye that is dispersed in the described substrate material; Described substrate material is for being the mixture that hydrophobic polymer, organo-siloxane and the organosilicon polymer of hydrocarbon chain forms by main chain; Described fluorescence dye is the rare earth compounding with photoluminescence performance.
2. fluorescent nano particles according to claim 1 is characterized in that: described main chain is that the hydrophobic polymer of hydrocarbon chain is one or more the miscellany of arbitrary combination in polymethacrylate, polyacrylic ester, polystyrene, methacrylic ester styrol copolymer, acrylate styrol copolymer, polymaleic acid dialkyl, dialkyl maleate styrol copolymer and the MBS.
3. fluorescent nano particles according to claim 1 is characterized in that: described organo-siloxane is RSi (OR ') 3, wherein R is the alkyl that 3～18 alkyl, thiazolinyl, phenyl or fluorine replace for the C carbonatoms, R ' is 1～2 alkyl for the C carbonatoms; Described organosilicon polymer is described organosilicone compounds RSi (OR ') 3Hydrolysis and the netted organosilicon polymer that forms by the condensation of silicon hydroxyl.
4. fluorescent nano particles according to claim 1, it is characterized in that: main chain is that hydrophobic polymer, the organo-siloxane of hydrocarbon chain, the mol ratio of rare earth compounding are 0.1～10: 0.005～3 in the described fluorescent nano particles: 1, and the quality percentage composition of organosilicon polymer is 0.5%～40% in the described fluorescent nano particles.
5. fluorescent nano particles according to claim 1 is characterized in that: described nanoparticle surface contains also that have can be for the active group of graft reaction.
6. fluorescent nano particles according to claim 4 is characterized in that: described active function groups for graft reaction is hydroxyl, amino, carboxyl or sulfydryl.
7. according to claim 1 or 2 or 3 or 4 or 5 described fluorescent nano particles, it is characterized in that: the skin of described nanoparticle also is enclosed with the silicon oxide shell.
8. fluorescent nano particles according to claim 7 is characterized in that: the thickness of described silicon oxide shell is 1～20nm.
9. according to claim 7 or 8 described fluorescent nano particles, the surface of described nanoparticle silicon oxide shell contains also that have can be for the active group of graft reaction, and described active function groups for graft reaction is hydroxyl, amino, carboxyl or sulfydryl.
10. according to fluorescent nano particles arbitrarily described in the claim 1-9, it is characterized in that: the particle diameter 10～200nm of described nanoparticle.
11. according to fluorescent nano particles arbitrarily described in the claim 1-9, it is characterized in that: the described rare earth compounding that is scattered in the nanoparticle-based material is the rare earth compounding with under visible light and/or near infrared light and/or ultraviolet excitation visible emitting or near infrared light performance.
12. fluorescent nano particles according to claim 11 is characterized in that: described rare earth compounding comprises europium complex and/or terbium coordination compound and/or ytterbium title complex and/or erbium title complex and/or neodymium title complex.
13. fluorescent nano particles according to claim 12 is characterized in that: described rare earth compounding is an europium complex; Described europium complex is that structural formula is the title complex shown in formula I or the formula II; Among formula I and the formula II, R 1, R 2Be that carbonatoms is 1 to 4 alkyl, R 3, R 4, R 5, R 6Be methyl or H, R 7, R 8Be methyl, three fluoro methyl or H;
Figure F2009102034073C00021
Formula I formula II.
14. the preparation method of the described fluorescent nano particles of claim 1 comprises the steps:
1) with rare earth compounding, main chain is the hydrophobic polymer of hydrocarbon chain and compound R Si (OR ') 3Be dissolved in can with the miscible organic solvent of water in, make that to contain concentration be 1.0 * 10 -6～1.0 * 10 -3The claim 11 of mol/L or 12 or 13 described rare earth compoundings, concentration is 1.0 * 10 -6～1.0 * 10 -2The main chain of mol/L is the hydrophobic polymer of hydrocarbon chain; Concentration is 1.0 * 10 -6～1.0 * 10 -2The compound R Si of mol/L (OR ') 3Solution;
2) under stirring or ultrasound condition, the solution that step 1) is obtained joins volume in its aqueous dispersant of 1～100 times, obtaining with the main chain is that the hydrophobic polymer, organo-siloxane of hydrocarbon chain and mixture that organosilicon polymer forms are substrate material, based on the colloidal sol of the fluorescent nano particles of rare earth compounding fluorescence dye;
Described can be one or more any mixing in methyl alcohol, ethanol, acetone and the tetrahydrofuran (THF) with the miscible organic solvent of water; Described dispersion agent is tensio-active agent, water-soluble polymer or water soluble protein; Described tensio-active agent is cationic, anionic, non-ionic type or amphoterics or their mixture; Described protein comprises serum albumin or ovalbumin; Described water-soluble polymer comprises PVP or PVA; Described main chain is that the hydrophobic polymer of hydrocarbon chain is one or more the miscellany of arbitrary combination in polymethacrylate, polyacrylic ester, polystyrene, methacrylic ester styrol copolymer, acrylate styrol copolymer, polymaleic acid dialkyl, dialkyl maleate styrol copolymer and the MBS; Described organo-siloxane is RSi (OR ") 3, R is the alkyl that 3～18 alkyl, thiazolinyl, phenyl or fluorine replace for the C carbonatoms, R ' is 1～2 alkyl for the C carbonatoms.
When dispersion agent is tensio-active agent or during water soluble organic polymer, its concentration of aqueous solution 1.0 * 10 -4～1.0 * 10 -2Mol/L; When dispersion agent was protein, its concentration of aqueous solution was 0.1～5.0mg/mL.
15. method according to claim 14 is characterized in that: described step 2), described aqueous dispersant is that concentration is 1.0 * 10 -4～1.0 * 10 -3The water phase surfactant mixture of mol/L, the volume ratio of the solution that described water phase surfactant mixture and described step 1) obtain is 1-50: 1.
16., it is characterized in that: comprise also in the described method that with concentration be 1.0 * 10 according to claim 14 or 15 described methods -6～1.0 * 10 -4The XRSi of mol/L (OR ") 3The alcohol or the aqueous solution join described step 2) in the colloidal sol of the fluorescent nano particles that obtains, make described RSi (OR ') 3With described XRSi (OR ") 3The usage quantity mol ratio be 1-200: 1, in 0～80 ℃ of reaction 1～24hr, making described surface contains and can be used for the active function groups of other compound covalent bonding, be that mixture that hydrophobic polymer, organo-siloxane and the organosilicon polymer of hydrocarbon chain forms is a substrate material with the main chain, based on the colloidal sol of the fluorescent nano particles of rare earth complex compoud; Described compounds X RSi (OR ") 3In, X is amino, NH 2CH 2CH 2NH-base, carboxyl or sulfydryl, XR be carbonatoms be 2～5 contain the substituent alkyl of X, OR " be methoxyl group, oxyethyl group, OH or ONa.
17. method according to claim 16 is characterized in that: described method is included in also in the described solution of step 1) that to add final concentration be 1.0 * 10 -6～1.0 * 10 -4The XRSi of mol/L (OR ") 3And in step 2) described this solution adds after the described water phase surfactant mixture, in 0～80 ℃ of reaction 1～24hr, making described surface contains and can be used for the active function groups of other compound covalent bonding, be that mixture that hydrophobic polymer, organo-siloxane and the organosilicon polymer of hydrocarbon chain forms is a substrate material with the main chain, based on the colloidal sol of the fluorescent nano particles of rare earth compounding; Described compounds X RSi (OR ") 3In, X is amino or sulfydryl, XR be carbonatoms be 2～5 contain the substituent alkyl of X, OR " be methoxyl group, oxyethyl group.
18. according to any described method among the claim 14-17, it is characterized in that: comprise also in the described method that with concentration be 5.0 * 10 -3Mol/L～5.0 * 10 -1The positive silicon ester alcoholic solution of mol/L joins in the colloidal sol of any fluorescent nano particles that obtains among the claim 14-17 according to 1: 5～100 volume ratio, in 0～80 ℃ of reaction 1～24hr, centrifugation, precipitation is scattered in the water, makes the colloidal sol of fluorescent nano particles with shell-core structure; Then, add 1.0 * 10 -6～1.0 * 10 -4The XRSi of mol/L (OR ") 3The alcohol or the aqueous solution, in 0～80 ℃ of reaction 1～24hr, make have shell-core structure, the surface contains the colloidal sol that can be used for the fluorescent nano particles of the active function groups of other compound covalent bondings; Wherein, the alcoholic solution of described positive silicon ester is the methyl alcohol or the ethanolic soln of methyl silicate or tetraethoxy; Described compounds X RSi (OR ") 3In, X is amino, NH 2CH 2CH 2NH-base, carboxyl or sulfydryl, XR be carbonatoms be 2～5 contain the substituent alkyl of X, OR " be methoxyl group, oxyethyl group, OH or ONa.
19. according to any described method among the claim 14-18, it is characterized in that: described main chain is the hydrophobic polymer of hydrocarbon chain and compound R Si (OR ') 3With the mol ratio of rare earth compounding be 0.1～10: 0.1～10: 1.
20. according to the method described in the claim 19, it is characterized in that: in the described method, comprise that also the fluorescent nano particles to obtaining carries out purifying, the method of described purifying is the centrifugal and collecting precipitation of colloidal sol of fluorescent nano particles that will obtain, to be deposited in the water then and disperse, perhaps prepared colloidal sol be carried out purifying with chromatography.
21. the application of any described fluorescent nano particles in synthesis of nano biological probe and fluoroimmunoassay among the claim 1-13.
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