CN102757789A - Rare earth doped gadolinium-potassium fluoride nanometer material for magneto-optical double-module biological marker and preparation method thereof - Google Patents
Rare earth doped gadolinium-potassium fluoride nanometer material for magneto-optical double-module biological marker and preparation method thereof Download PDFInfo
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Abstract
The invention relates to a rare earth doped gadolinium-potassium fluoride nanometer material for a magneto-optical double-module biological marker and a preparation method thereof. The preparation method comprises the following steps of: mixing gadolinium chloride, potassium chloride and ammonium fluoride in distilled water, ethanol and ethanediol by utilizing propylene imine as a surface active agent; carrying out hydro-thermal treatment at 50-230 DEG C for a period of time; washing, and drying to obtain a DGdF4:Ln nanometer crystal, wherein the commonest of the DGdF4:Ln nanometer crystal are as follows: xLn3+-(1-x)KGdF4 (Ln=Ce, Pr, Nd, Pm, Sm, Eu, Tb, Dy, Ho, Er, Tm, Yb; x=0-60 mol%). The rare earth doped DGdF4 nanometer fluorescence marker material prepared by adopting the method can not only control the size of nanometer granules at about 25 nanometers, but also has better water solubility, can utilize the amino on the surface to be connected with biological molecules; besides, the biological connection is subjected to hypersensitivity detection by realizing needed specific fluorescence emission in such a way that different rare earth ions are doped in the nanometer granules, namely the rare earth doped DGdF4 nanometer fluorescence marker material obtained through the preparation method has potential in being applied in the field of biological markers; and because gadolinium ions are contained in a matrix, the DGdF4 nanometer crystal can be further used as a T1 magnetic resonance imaging contrast agent.
Description
Technical field
The present invention relates to magneto-optic bimodulus biological labled material and preparation method thereof, especially relate to the preparation of rear-earth-doped gadolinium fluoride potassium nano material and in the application in magneto-optic bimodulus biomarker field.
Background technology
The multimode biological detection because its imaging that can realize light, nuclear, magnetic, various modes such as ultrasonic simultaneously detects, and received scientific circles and society's concern more and more widely, and wherein magneto-optic bimodulus biomarker is particularly noticeable.This is owing to can provide the light of detection sensitivity the highest in the detection means to detect the restriction that receives investigation depth easily, detects though nuclear magnetic resonance can be carried out three-dimensional, has limited its widespread use because of lower detectivity and resolving power.The magneto-optic bimodulus biomarker that therefore can two kinds of detecting patterns of magneto-optic be concentrated in together just can be realized detection in the body of high resolving power, highly sensitive and living microbe.Current magneto-optic bimodulus marker material be mostly through ultra paramagnetic ferric oxide nanometer particle and organic dye or quantum dot combine prepare.Yet this hybrid material but faces many difficulties in the application in biomarker field, like ultra paramagnetic iron oxide nano particle the magnetic inductive effect of normal cell tissue is unfavorable for that its nuclear magnetic resonance in vivo detects; Weakness such as the photobleaching of organic dye, emission peak width, photochromism are impure cause the instability of its detectable signal; Quantum dot toxicity is bigger, and preparation cost is higher, and is unsuitable to biological in-vivo imaging marker material.In addition, two kinds of different particles are combined, tend to destroy their optics and magnetism characteristics separately, and particle size is increased, thereby cause effect to be had a greatly reduced quality.（Kim,?J.?et?al.?“Multifunctional?Uniform?Nanoparticles?Composed?of?a?Magnetite?Nanocrystal?Core?and?a?Mesoporous?Silica?Shell?for?Magnetic?Resonance?and?Fluorescence?Imaging?and?for?Drug?Delivery”?Angewandte?Chemie-International?Edition?Vol.47,?8438-8441,?2008；Huh,?Y.?M.?et?al.?“In?vivo?magnetic?resonance?detection?of?cancer?by?using?multifunctional?magnetic?nanocrystals”?Journal?of?the?American?Chemical?Society?Vol.127,?12387-12391,?2005）。Rear-earth-doped gadolinium-contained nano material, because its good luminous and magnetic performance, and extensively thought the magneto-optic bimodulus biological labled material that a new generation is best.Rear-earth-doped nanocrystalline fluorescence emission peak is narrower, colourity is pure, the life-span is long, light is stable, relatively is suitable as the biological fluorescent labelling material.Gadolinium ion is considered to T best in the periodic table of elements because its 4f electronic shell contains seven single electrons
1The metals ion of magnetic resonance imaging contrast.What is more important and since luminous rare earth ion with provide the gadolinium ion of magnetic inductive effect to come from same nano particle, so this method can avoid complicated synthesis program to become greatly with particle size.Fluorochemical, particularly AREF
4The fluorochemical of (A=basic metal, RE=rare earth ion) structure is because its phonon energy is low and can realize the high-concentration dopant of other rare earth ions, so be widely used as rare earth ion doped matrix.The present invention can realize that through the different rare earth ion that in gadolinium fluoride potassium nano material, mixes it is multicolor luminous.The amino that utilizes nano grain surface simultaneously can be connected with biomolecules and be applied to biomarker.And utilizing the gadolinium ion that contains in the matrix, this nano particle can carry out T
1The nuclear magnetic resonance mark.
Summary of the invention
The present invention proposes a kind of rear-earth-doped gadolinium fluoride potassium preparation of nanomaterials that is used for magneto-optic bimodulus mark.
The component of the rear-earth-doped gadolinium fluoride potassium nano material of the present invention's preparation is: xLn
3+-(1-x) KGdF
4(Ln=Ce, Pr, Nd, Sm, Eu, Tb, Dy, Ho, Er, Tm, Yb; X=0-60 mol%).
The present invention adopts and is prepared as follows technology: rare-earth chlorination salt, Gadolinium trichloride, Repone K and Vestolen PP 7052 imines are dissolved in zero(ppm) water, ethanol and the terepthaloyl moietie fully; Other is dissolved in an amount of Neutral ammonium fluoride in zero(ppm) water and the ethanol and is added drop-wise to above-mentioned solution, then 50-230 ℃ of following hydrothermal treatment consists.Can obtain the KGdF of surface amination after the separation of deposition process, washing, the drying
4: the Ln manocrystalline powders.
The sign of described rear-earth-doped gadolinium fluoride potassium nano material: detect through X-ray powder diffraction (XRD) experiment and to show that the gadolinium fluoride potassium of preparing is nanocrystalline and be pure cube of phase structure.What transmission electron microscope (TEM) test demonstration obtained is the homodisperse nano particle about 25 nanometers; High-resolution-ration transmission electric-lens (HRTEM) photo can clearly be observed even successive lattice fringe, shows that the nano particle crystallization is better.In addition, in nanocrystalline, can realize needed specific fluorescent emission through the different rare earth ion that mixes.
Selecting for use according to propyleneimine is tensio-active agent, and both the nanocrystalline size of may command can realize the amino functional that it is surperficial again.
The purposes of resultant amidized rear-earth-doped gadolinium fluoride potassium nano material is characterized in that: be applied to magneto-optic bimodulus biomarker and detection.The rear-earth-doped gadolinium fluoride potassium nano material of preparation gained utilizes its surperficial amino to be connected with biomolecules; Simultaneously can be through specific luminous of the rare earth ion realization of mixing different in the rear-earth-doped gadolinium fluoride potassium nano material; Come this biological connection is detected, be applied to fields such as biological mark.
Material preparation process of the present invention is simple, cost is low, good reproducibility, can produce in enormous quantities.The present invention compares with other magneto-optic bimodulus biological labled materials that prepare both at home and abroad at present, the nano-crystalline granule homogeneous, and size is little, and emission peak is narrow, and luminous intensity is high, and luminescent lifetime is long, and magnetic relaxation rate is big, is a kind of good novel magneto-optic bimodulus biological labled material.Nano grain surface has amino group, has better water solubility, can directly should make magneto-optic bimodulus biological labled material.
Description of drawings
Accompanying drawing 1:KGdF
4: the nanocrystalline synoptic diagram of Ln as magneto-optic bimodulus marker material.
Accompanying drawing 2:KGdF
4: Eu
3+(2.5 mol%) nanocrystalline XRD diffraction pattern.
Accompanying drawing 3: mix KGdF
4: Tb
3+(2.5 mol%) nanocrystalline transmission electron microscope picture.
Accompanying drawing 4: mix KGdF
4: Dy
3+The high-resolution-ration transmission electric-lens figure that (2.5 mol%) is nanocrystalline.
Accompanying drawing 5: the KGdF of embryo lung fibroblast and different concns
4The activity figure of nanocrystalline cultivation after 24 hours.
Accompanying drawing 6: mix Ln
3+1.5at% (Ln=Eu
3+, Tb
3+, Dy
3+) KGdF
4Nanocrystalline excites and emmission spectrum.
Accompanying drawing 7:KGdF
4: Tb
3+Nanocrystalline biotinylated schematic flow sheet.
Accompanying drawing 8: biotinylated KGdF
4: Tb
3+(2.5 mol%) is for the detection curve of the avidin of different concns.
Accompanying drawing 9:KGdF
4Nanocrystalline (a) mr relaxation rate and (b) the nuclear magnetic resonance synoptic diagram of the different concns aqueous solution.
Embodiment
Instance 1: take by weighing 0.0037 g Europium trichloride (EuCl
36H
2O), 0.8 g Repone K (KCl) and 0.37 g Gadolinium trichloride (GdCl
36H
2O) be dissolved in fully in the 12 mL zero(ppm) water, add 1 mL ethanol, 20 mL terepthaloyl moietie and 10 mL Vestolen PP 7052 imide liquors then, stir and obtain clear solution A; In another beaker, an amount of Neutral ammonium fluoride is dissolved in the 0.5 mL zero(ppm) water, adds 2.5 mL ethanol then, stir and obtain clear solution B; Under whipped state, in solution B, dropwise join solution A, continue to stir ten minutes, solution is transferred in the hydro-thermal jar, 170 ℃ of following hydrothermal treatment consists 20 hours.After the cooling, supernatant liquid is removed, lower sediment water and ethanol are washed after drying in turn three times, drying obtains KGdF under 60 ℃ of vacuum conditions then
4: Eu
3+Manocrystalline powders.This manocrystalline powders is dissolved in zero(ppm) water, can obtains colourless transparent solution, excite down, can observe strong emitting red light at 272.5 nm.
Instance 2: take by weighing 0.0037g terbium chloride (TbCl
36H
2O), 1.8g Repone K (KCl) and 0.37g Gadolinium trichloride (GdCl
36H
2O) be dissolved in fully in the 5mL zero(ppm) water, add 25mL ethanol, 5mL terepthaloyl moietie and 5 mL Vestolen PP 7052 imide liquors then, stir and obtain clear solution A; In another beaker, an amount of Neutral ammonium fluoride is dissolved in the 1mL zero(ppm) water, adds 1mL ethanol then, stir and obtain clear solution B; Under whipped state; In solution B, dropwise join solution A, continue to stir ten minutes, solution is transferred in the hydro-thermal jar; 110 ℃ of following hydrothermal treatment consists 18 hours. after the cooling; Supernatant liquid is removed, lower sediment water and ethanol are washed after drying in turn three times, drying obtains KGdF under 50 ℃ of vacuum conditions then
4: the Tb manocrystalline powders.With 2 milligrams of KGdF
4: Tb
3+Manocrystalline powders is dissolved in the zero(ppm) water of 20mL, can obtain KGdF
4: the Tb colourless transparent solution.Excite down with 272.5 nm, can see the luminous of green.
Instance 3: take by weighing 0.0037g Dysprosium trichloride (DyCl
36H
2O), 1.8g Repone K (KCl) and 0.37g Gadolinium trichloride (GdCl
36H
2O) be dissolved in fully in the 2mL zero(ppm) water, add 28mL terepthaloyl moietie and 0.5 mL Vestolen PP 7052 imide liquor then, stir and obtain clear solution A; In another beaker, an amount of Neutral ammonium fluoride is dissolved in the 5mL zero(ppm) water, adds 1mL ethanol then, stir and obtain clear solution B; Under whipped state; In solution B, dropwise join solution A, continue to stir ten minutes, solution is transferred in the hydro-thermal jar; 140 ℃ of following hydrothermal treatment consists 24 hours. after the cooling; Supernatant liquid is removed, lower sediment water and ethanol are washed after drying in turn three times, drying obtains KGdF under 65 ℃ of vacuum conditions then
4: Dy
3+Manocrystalline powders.With 15 milligrams of KGdF
4: the Dy manocrystalline powders is dissolved in the zero(ppm) water of 20mL, can obtain KGdF
4: the Dy colourless transparent solution.Excite down with 272.5nm, can see the luminous of blueness.
Instance 4: the KGdF that takes by weighing different mass
4Nanocrystalline, join respectively in the petridish of embryo lung fibroblast of equal amts, at 37 degrees centigrade, 5%CO
2Incubator in hatch 24 hours after, utilize the cytoactive detection technique, find the KGdF of homeomorphism lung fibroblast not in big concentration
4In the nanocrystalline aqueous solution, still keep the activity more than 95%, KGdF is described
4Nanocrystalline have a bio-compatibility.
Instance 5:KGdF
4: the preparation of the nanocrystalline connection vitamin H of Tb.Take by weighing 0.2 mmol vitamin H (biotin) and 0.2 mmol 1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimide (EDC) and 0.5 mmol N-hydroxy-succinamide (NHS); Be dissolved in 20 mL phosphate buffered saline buffers (pH=7.2); Stirred 15 minutes, then to wherein adding 20 mg KGdF
4: the Tb nano particle, stirred 1 day, with zero(ppm) water and washing with alcohol repeatedly dry in vacuum drying oven then with precipitation and centrifugal separation, can obtain the KGdF of finishing vitamin H
4: the Tb inorganic nano-crystal.
Instance 6: the KGdF that will connect vitamin H
4: the nanocrystalline phosphate buffered saline buffer (pH=7.2) that is dissolved in of Tb; Be divided into several parts of equal volume; After cultivating with the avidin that is marked with fluorescein isothiocyanate (FITC-avidin) of different concns then; On the fluorescence plate reading machine, take time resolved spectroscopy to detect, excitation wavelength is 253 nm, and be 100 time of lag
μS, the gate time is 1 ms.Because Doped Rare Earth ion Tb in the nano particle
3+With the fluorescence resonance energy transmission (FRET) that resorcinolphthalein FITC exists, the result shows along with the increase of the FITC-avidin concentration that adds, detects Tb
3+Luminous intensity corresponding to 489 nm wave bands reduces; Resorcinolphthalein FITC increases corresponding to the luminous intensity of 520 nm wave bands simultaneously; 3.7-200 (?) in the nM scope; Exist the better linearity relation between avidin concentration and fluorescence intensity change, explain that the fluorescence that utilizes nano particle can be applied to detect avidin.
Instance 7: with KGdF
4Nanocrystalline being dissolved in the water, and be diluted to the KGdF of different concns
4The aqueous solution.At medical magnetic resonance imager T
1Detect under the sequence, along with KGdF
4The increase of concentration, the picture brightness that obtains increases gradually, and KGdF is described
4Can be used as T
1Magnetic resonance imaging contrast.
Claims (3)
1. rear-earth-doped gadolinium fluoride potassium preparation of nanomaterials that is used for magneto-optic bimodulus biomarker, its component is: the xLn of surface amination
3+-(1-x) KGdF
4, Ln=Ce wherein, Pr, Nd, Sm, Eu, Tb, Dy, Ho, Er, Tm, Yb; X=0-60mol%.
2. rear-earth-doped gadolinium fluoride potassium preparation of nanomaterials according to claim 1 is characterized in that: under the room temperature, rare-earth chlorination salt, Repone K, Gadolinium trichloride are dissolved in fully in the aqueous solution of zero(ppm) water, ethanol, terepthaloyl moietie and Vestolen PP 7052 imines; In addition an amount of Neutral ammonium fluoride is dissolved in zero(ppm) water and the terepthaloyl moietie, and dropwise adds in the above-mentioned solution, carry out hydrothermal treatment consists at 50-230 ℃ then, at last with precipitate and separate, washing, drying.
3. the purposes of rear-earth-doped gadolinium fluoride potassium nano material according to claim 1 is characterized in that: this material can be used as magnetic resonance imaging contrast; Utilize the amino of this material surface of preparation gained to be connected with biomolecules, Doped Rare Earth ionic capable of using is specific luminous simultaneously, comes this biological connection is detected, and then can be applicable to field such as biological fluorescent labelling.
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Cited By (4)
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| CN102925155A (en) * | 2012-11-05 | 2013-02-13 | 昆明理工大学 | Near infrared fluorescent probe substrate material of rare earth ion nano alkali metal rare earth fluoride and preparation method of near infrared fluorescent probe substrate material |
| CN103275722A (en) * | 2013-06-20 | 2013-09-04 | 南京大学 | Magneto-optical bimodal imaging probe rare earth nanoparticle, and preparation method and application thereof |
| CN106978179A (en) * | 2017-03-31 | 2017-07-25 | 武汉理工大学 | A kind of 808nm is excited, Li+Adulterate KGdF4：Nd3+Nano luminescent material and preparation method thereof |
| CN107903899A (en) * | 2017-12-21 | 2018-04-13 | 内蒙古师范大学 | A kind of ytterbium erbium is co-doped with KGdF4The synthetic method of nano particle and nanobelt |
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| CN101205461A (en) * | 2007-11-23 | 2008-06-25 | 浙江大学 | In-situ surface finishing method for lanthanide doped fluoride luminescent nano particles |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102925155A (en) * | 2012-11-05 | 2013-02-13 | 昆明理工大学 | Near infrared fluorescent probe substrate material of rare earth ion nano alkali metal rare earth fluoride and preparation method of near infrared fluorescent probe substrate material |
| CN102925155B (en) * | 2012-11-05 | 2014-01-15 | 昆明理工大学 | Near infrared fluorescent probe substrate material of rare earth ion nano alkali metal rare earth fluoride and preparation method of near infrared fluorescent probe substrate material |
| CN103275722A (en) * | 2013-06-20 | 2013-09-04 | 南京大学 | Magneto-optical bimodal imaging probe rare earth nanoparticle, and preparation method and application thereof |
| CN103275722B (en) * | 2013-06-20 | 2016-08-10 | 南京大学 | A kind of magneto-optic bimodal image probe rare earth nanoparticle and preparation method thereof and purposes |
| CN106978179A (en) * | 2017-03-31 | 2017-07-25 | 武汉理工大学 | A kind of 808nm is excited, Li+Adulterate KGdF4：Nd3+Nano luminescent material and preparation method thereof |
| CN107903899A (en) * | 2017-12-21 | 2018-04-13 | 内蒙古师范大学 | A kind of ytterbium erbium is co-doped with KGdF4The synthetic method of nano particle and nanobelt |
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