CN1524925A - Fluorescent magnetic nanometer compound, preparing process and application thereof - Google Patents
Fluorescent magnetic nanometer compound, preparing process and application thereof Download PDFInfo
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- CN1524925A CN1524925A CNA031271650A CN03127165A CN1524925A CN 1524925 A CN1524925 A CN 1524925A CN A031271650 A CNA031271650 A CN A031271650A CN 03127165 A CN03127165 A CN 03127165A CN 1524925 A CN1524925 A CN 1524925A
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- fluorescence
- magnetic nano
- particle
- polyelectrolyte
- absorption
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Abstract
The invention relates to a fluorescent magnetic nano compound, its preparation method and use, belonging to the field of nano material, Wherein single magnetic nano particle is used as a core with coated polyelectrolyte multilayers from layer to layer, magnetic nano particles, polyelectrolyte multilayers, semi-conductor fluorescent nano crystal and polyelectrolyte multilayers, the polyelectrolyte multilayers are the 1-30 layers coated at intervals by poly positive ionic electrolyte and poly negative ionic electrolyte. And its preparation process comprises the steps of adsorbing polyelectrolyte, adsorbing magnetic nano particles, and adsorbing semi-conductor fluorescent nano crystal.
Description
Technical field
The invention belongs to field of nanometer material technology, particularly fluorescence, magnetic nanocomposites and preparation method thereof, and be coupled by control surface character and biomolecule are applied to separation, the orientation of biological sample and detect.
Background technology
In recent years, semiconductor fluorescence is nanocrystalline because its special physics and chemical property have begun to be widely used in fields such as optics, biology, medical science as fluorescent marker of new generation.Utilize quantum size effect and the quantum confined effect of fluorescence nano, can obtain having simply the material of different glow colors by changing particle diameter.Utilize the continuity of fluorescence nano excitation spectrum and the narrow and symmetric characteristic of emmission spectrum, cross interference in the time of can significantly reducing the hyperchannel fluoroscopic examination, make that selecting for use a kind of excitation wavelength to carry out the polychrome analysis becomes possibility, thereby realize multi-color marking and the joint inspection of many indexs.The characteristics of luminescence of the efficient stable of fluorescence nano and long fluorescence lifetime also help improving the sensitivity of detection, have enlarged the kind of detectable disease, have realized the quantification that detects.Calendar year 2001, it is in 1.2 microns the polystyrene microsphere that the people such as Nie of U.S. Indiana University will have the nanocrystalline diameter that is incorporated into of the CdSe of different colours and luminous intensity, thereby has realized the multicolor fluorescence coding.When the fluorescence nano of selecting 6 kinds of different glow colors and 10 kinds of different luminous intensities carries out any combination, encode just can in theory 1,000,000 protein moleculars.
The advantage of this method is to only depend on simple low-speed centrifugal just can successfully product separation be come out, shortcoming is owing to utilized the swelling characteristic of polystyrene microsphere in special solvent, make fluorescence nano enter in the microballoon, be subjected to the influence of environment bigger, easily cause the leakage of fluorescence nano; And owing to make carrier with polymer microsphere, the particle diameter of product is bigger, is generally submicron to micrometer range, thereby has limited the range of application of product greatly.If adopt SiO
2Coat the method for fluorescence nano, although end product is more stable, because the particle diameter of product has been decreased to nanometer scale, brought new problem for again separation and purification.
Summary of the invention
The technical problem to be solved in the present invention is a kind of fluorescence magnetic nano-complex of design, has little, the good stability of granularity, can be coupled with the antibody class biological sample, can realize that not only the multicolor fluorescence of antagonist class biological sample detects, can realize simultaneously again separation, orientation and enrichment to product; Design a kind of method for preparing this fluorescence magnetic nano-complex, and device is simple, saves cost, time saving and energy saving, easy to operate.
Fluorescence magnetic nano-complex of the present invention is to be nuclear with single magnetic nano-particle, and nuclear is outer successively to coat polyelectrolyte multilayer film, magnetic nano-particle and polyelectrolyte multilayer film, semiconductor fluorescence is nanocrystalline and polyelectrolyte multilayer film; Its granularity is controlled between 10～300nm; Said polyelectrolyte multilayer film is polycation electrolyte and the alternate coating of polyanion electrolyte 1～30 layer.
The magnetic intensity of fluorescence magnetic nano-complex increases with the increase of the amount of the magnetic particle of absorption, can increase by 1～100 times than the magnetic intensity of single magnetic nano particle daughter nucleus; Its fluorescence intensity also can increase with the increase of the semiconductor nano number of plies of adsorbing.
Fluorescence magnetic nano-complex of the present invention adopts LBL (layer-by-layer) technology by polycation electrolyte/polyanion electrolyte layer fluorescence nano to be assembled on the magnetic particle.
Concrete preparation method is, with magnetic nano-particle, polyelectrolyte, semiconductor fluorescence is nanocrystalline is raw material, control magnetic nano-particle, polyelectrolyte, the nanocrystalline mol ratio of semiconductor fluorescence are 1: (15～6250): (20～100); Experience absorption polyelectrolyte---absorption magnetic nano-particle---the nanocrystalline technological process of absorption semiconductor fluorescence; Said absorption polyelectrolyte is to be nuclear with single magnetic nano-particle, in salt concn is that 0.5～20mmol/L, pH value are under 8～10 conditions, at the alternate absorption polyelectrolyte multilayer film in magnetic nano particle daughter nucleus surface, every absorption one deck just utilizes remaining polyelectrolyte and impurity in the magnetic separation technique flush away system; Said absorption magnetic nano-particle, be in magnetic nano particle daughter nucleus surface adsorption behind 1～30 strata ionogen, alternate absorption magnetic nano-particle and polyelectrolyte multilayer film, the quantity of the magnetic nano-particle of absorption is 10～100 times of single magnetic nano particle daughter nucleus, every absorption one deck just utilizes magnetic separation technique to wash, and forms the magnetic Nano mixture; Said absorption semiconductor fluorescence is nanocrystalline to be, it is nanocrystalline to add semiconductor fluorescence in the magnetic Nano complex solution, utilize the magnetic separation technique washing, adsorb polyelectrolyte again, adsorb the nanocrystalline and polyelectrolyte of semiconductor fluorescence through layer by layer, obtain the fluorescence magnetic nano-complex, the nanocrystalline each adsorptive capacity of semiconductor fluorescence is previous 1～2 times; Said polyelectrolyte multilayer film is polycation electrolyte and the alternate coating of polyanion electrolyte 1～30 layer.
Aforesaid magnetic nano-particle can be selected Fe
3O
4, Fe
2O
3, CoFe
2O
4, NiFeO
4, ZnFe
2O
4, MnFeO
4Deng; The nanocrystalline optional CdS of semiconductor fluorescence, CdSe, CdTe, ZnS etc. and their doped and compounded thing thereof.Salt in the aforesaid absorption polyelectrolyte body can be sodium-chlor (NaCl), saltpetre (KNO
3) etc.
The application of fluorescence magnetic nano-complex of the present invention in the orientation of the separation of the detection of biological sample, biological sample and biological sample at first adds biological sample in the fluorescence magnetic nanocomposite system, be coupled under 0～10 ℃ of condition; Carrying out afterwards the biological sample multicolor fluorescence detects or the Magnetic Isolation of biological sample or the magnetic orientation of biological sample.Biological sample (antibody) concentration can be at 30～100 μ g/ml, coupling connection under 3～5 ℃ of conditions.Here said biological sample comprises protein, amino acid, DNA, antigen, antibody etc.
The present invention can realize that magnetic property, photoluminescent property, size and the surface properties height of fluorescence magnetic nano-complex is controlled.That is, can prepare the fluorescence magnetic nano-complex of magnetic property not of the same race, photoluminescent property, size and surface properties.The magnetic power can realize by the magnetic material of selecting variety classes or different-grain diameter, and the number of plies of magnetic particle that also can be by assembling is controlled.Photoluminescent property mainly comprises fluorescence intensity and emission wavelength, can realize by the fluorescence nano of selecting variety classes or different-grain diameter, and the number of plies of the fluorescence nano that fluorescence intensity also can be by assembling is controlled.The size of nano-complex can be controlled by the electrolyte of assembling and the number of plies of nano particle.The surface properties of fluorescence magnetic nano-complex can realize by the surface properties of fluorescence nano, the kind of polyelectrolyte, further functionalisation of surfaces (silanization or surface aggregate polymer).
Preparation method's device of fluorescence magnetic nano-complex of the present invention is simple, easy to operate; Utilize the magnetic property of magnetic particle, under externally-applied magnetic field, can realize being labeled the separation of thing; Time saving and energy saving, save cost.The fluorescence magnetic nano-complex product that makes can realize that not only multicolor fluorescence detects, and can realize simultaneously again separation, orientation and the enrichment of product, can be widely used in a plurality of fields such as biochemistry, molecular biology, clinical medicine.
Embodiment
Embodiment 1
(1). magnetic nanocomposites [Fe
3O
4/ (polycation electrolyte/polyanion electrolyte)
m]
x[be called for short (Fe
3O
4/ PE
m)
x] synthetic.
Be that to join 1ml, concentration be that 25mg/ml, pH value are 9, contain the Fe of 20mM NaCl to 10mg/ml polycation electrolyte (containing 20mM NaCl) 100 μ l with concentration
3O
4In the solution, reacted 20 minutes, utilize magnetic separation technique washing after, the pH value that again be scattered in 1ml, contains 20mM NaCl is in 9 the cushioning liquid.To add 100 μ l concentration again be 10mg/ml, contain the polyanion electrolyte of 20mM NaCl, react after 20 minutes, the magnetic resolution washing, be scattered in 1ml again, contain 20mM NaCl, the pH value is in 9 the buffered soln.Repeat above-mentioned steps, after particle surface has adsorbed m strata electrolyte, add again 50mlFe
3O
4, after the Magnetic Isolation washing, repeat x above-mentioned whole step (x=1～30), obtain shell thickness with the strong and weak different magnetic nanocomposites (Fe of the magnetic that increase increases, positive charge is rich on the surface (or negative electrical charge) of the absorption number of plies
3O
4/ PE
m)
x
Wherein, magnetic intensity increases with the increase of the magnetic nano-particle number of plies of absorption, and the increase scope can be 10～100 times of stratum nucleare magnetic nano-particle.
(2). have the fluorescence magnetic nano-complex (Fe of different fluorescence intensities and emission wavelength
3O
4/ PE
m)
x/ (CdTe/PE
n)
ySynthetic.
To add 5ml, concentration and be 3mM, pH value in the product of (1) be 9, contain the CdTe solution of 20mM NaCl, reacts after 20 minutes, after the magnetic resolution washing, be scattered in 1ml, contain 20mM NaCl, the pH value is in 9 the buffered soln.Adding 100 μ l concentration is 10mg/ml polycation electrolyte (containing 20mMNaCl) again, reacted 20 minutes, the Magnetic Isolation washing, repeat above-mentioned steps, after particle surface has adsorbed n strata electrolyte, add again CdTe, behind the separating, washing, repeat y above-mentioned whole step (y=1～30), finally can obtain (Fe
3O
4/ PE
m)
x/ (CdTe/PE
n)
yThe fluorescence magnetic nano-complex.Wherein, fluorescence intensity can increase by 1～1000 times with the increase of the semiconductor nano amount of absorption.Emission wavelength red shift clocklike along with the increase of CdTe particle diameter can be in the interval adjustment of 500～730nm.
The biological coupling connection of embodiment 2 fluorescence magnetic nano-complexes.
The antibody that in the fluorescence magnetic nano-complex that embodiment 1 makes, adds 5mg/ml, 4 ℃ of reactions one hour, 4 ℃ of centrifuge washings three times disperse again, and preserve under 4 ℃ of conditions.
Embodiment 3:
With the Fe among the embodiment 1
3O
4Be changed to Fe
2O
3, other conditionally complete is consistent, obtains (the Fe that shell thickness and fluorescence intensity increase with absorption number of plies x, y
2O
3/ PE
m)
x/ (CdTe/PE
n)
yThe fluorescence magnetic nano-complex.Also can obtain corresponding biology by embodiment 2 and be coupled product.
Embodiment 4:
With the Fe among the embodiment 1
3O
4Be changed to CoFe
2O
4, other conditionally complete is consistent, obtains (the CoFe that shell thickness and fluorescence intensity increase with absorption number of plies x, y
2O
4/ PE
m)
x/ (CdTe/PE
n)
yThe fluorescence magnetic nano-complex.Also can obtain corresponding biology by embodiment 2 and be coupled product.
Embodiment 5:
With the Fe among the embodiment 1
3O
4Be changed to NiFeO
4, other conditionally complete is consistent, obtains (the NiFe that shell thickness and fluorescence intensity increase with absorption number of plies x, y
2O
4/ PE
m)
x/ (CdTe/PE
n)
yThe fluorescence magnetic nano-complex.Also can obtain corresponding biology by embodiment 2 and be coupled product.
Embodiment 6:
With the Fe among the embodiment 1
3O
4Be changed to ZnFe
2O
4, other conditionally complete is consistent, obtains (the ZnFe that shell thickness and fluorescence intensity increase with absorption number of plies x, y
2O
4/ PE
m)
x/ (CdTe/PE
n)
yThe fluorescence magnetic nano-complex.Also can obtain corresponding biology by embodiment 2 and be coupled product.
Embodiment 7:
With the Fe among the embodiment 1
3O
4Be changed to MnFeO
4, other conditionally complete is consistent, obtains (the MnFe that shell thickness and fluorescence intensity increase with absorption number of plies x, y
2O
4/ PE
m)
x/ (CdTe/PE
n)
yThe fluorescence magnetic nano-complex.Also can obtain corresponding biology by embodiment 2 and be coupled product.
Embodiment 8:
CdTe among the embodiment 1 is changed to CdSe, and other conditionally complete is consistent, obtains (the Fe that shell thickness and fluorescence intensity increase with absorption number of plies x, y
3O
4/ PE
m)
x/ (CdSe/PE
n)
yThe fluorescence magnetic nano-complex.Also can obtain corresponding biology by embodiment 2 and be coupled product.
Embodiment 9:
CdTe among the embodiment 1 is changed to ZnS, and other conditionally complete is consistent, obtains (the Fe that shell thickness and fluorescence intensity increase with absorption number of plies x, y
3O
4/ PE
m)
x/ (ZnS/PE
n)
yThe fluorescence magnetic nano-complex.Also can obtain corresponding biology by embodiment 2 and be coupled product.
Embodiment 10:
CdTe among the embodiment 1 is changed to CdSe, and other conditionally complete is consistent, obtains (the Fe that shell thickness and fluorescence intensity increase with absorption number of plies x, y
2O
3/ PE
m)
x/ (CdSe/PE
n)
yThe fluorescence magnetic nano-complex.Also can obtain corresponding biology by embodiment 2 and be coupled product.
Embodiment 11:
CdTe among the embodiment 1 is changed to ZnS, and other conditionally complete is consistent, obtains (the Fe that shell thickness and fluorescence intensity increase with absorption number of plies x, y
2O
3/ PE
m)
x/ (ZnS/PE
n)
yThe fluorescence magnetic nano-complex.Also can obtain corresponding biology by embodiment 2 and be coupled product.
Embodiment 12:
CdTe among the embodiment 4 is changed to CdSe, and other conditionally complete is consistent, obtains (the CoFe that shell thickness and fluorescence intensity increase with absorption number of plies x, y
2O
4/ PE
m)
x/ (CdSe/PE
m)
yThe fluorescence magnetic nano-complex.Also can obtain corresponding biology by embodiment 2 and be coupled product.
Embodiment 13:
CdTe among the embodiment 4 is changed to ZnS, and other conditionally complete is consistent, obtains (the CoFe that shell thickness and fluorescence intensity increase with absorption number of plies x, y
2O
4/ PE
m)
x/ (ZnS/PE
n)
yThe fluorescence magnetic nano-complex.Also can obtain corresponding biology by embodiment 2 and be coupled product.
Embodiment 14:
CdTe among the embodiment 5 is changed to CdSe, and other conditionally complete is consistent, obtains (the NiFe that shell thickness and fluorescence intensity increase with absorption number of plies x, y
2O
4/ PE
m)
x/ (CdSe/PE
n)
yThe fluorescence magnetic nano-complex.Also can obtain corresponding biology by embodiment 2 and be coupled product.
Embodiment 15:
CdTe among the embodiment 5 is changed to ZnS, and other conditionally complete is consistent, obtains (the NiFe that shell thickness and fluorescence intensity increase with absorption number of plies x, y
2O
4/ PE
m)
x/ (ZnS/PE
n)
yThe fluorescence magnetic nano-complex.Also can obtain corresponding biology by embodiment 2 and be coupled product.
Embodiment 16:
CdTe among the embodiment 6 is changed to CdSe, and other conditionally complete is consistent, obtains (the ZnFe that shell thickness and fluorescence intensity increase with absorption number of plies x, y
2O
4/ PE
m)
x/ (CdSe/PE
n)
yThe fluorescence magnetic nano-complex.Also can obtain corresponding biology by embodiment 2 and be coupled product.
Embodiment 17:
CdTe among the embodiment 6 is changed to ZnS, and other conditionally complete is consistent, obtains (the ZnFe that shell thickness and fluorescence intensity increase with absorption number of plies x, y
2O
4/ PE
m)
x/ (ZnS/PE
n)
yThe fluorescence magnetic nano-complex.Also can obtain corresponding biology by embodiment 2 and be coupled product.
Claims (4)
1, a kind of fluorescence magnetic nano-complex is characterized in that, is nuclear with single magnetic nano-particle, and nuclear successively coats polyelectrolyte multilayer film, magnetic nano-particle and polyelectrolyte multilayer film outward, semiconductor fluorescence is nanocrystalline and polyelectrolyte multilayer film; Its granularity is in 10～300nm scope; Said polyelectrolyte multilayer film is polycation electrolyte and the alternate coating of polyanion electrolyte 1～30 layer.
2, a kind of preparation method of fluorescence magnetic nano-complex of claim 1, it is characterized in that, with magnetic nano-particle, polyelectrolyte, semiconductor fluorescence is nanocrystalline is raw material, control magnetic nano-particle, polyelectrolyte, the nanocrystalline mol ratio of semiconductor fluorescence are 1: (15～6250): (20～100); Experience absorption polyelectrolyte---absorption magnetic nano-particle---the nanocrystalline technological process of absorption semiconductor fluorescence; Said absorption polyelectrolyte is to be nuclear with single magnetic nano-particle, in salt concn is that 0.5～20mmol/L, pH value are under 8～10 conditions, at the alternate absorption polyelectrolyte multilayer film in magnetic nano particle daughter nucleus surface, every absorption one deck just utilizes remaining polyelectrolyte and impurity in the magnetic separation technique flush away system; Said absorption magnetic nano-particle, be in magnetic nano particle daughter nucleus surface adsorption behind 1～30 strata ionogen, alternate absorption magnetic nano-particle and polyelectrolyte multilayer film, the quantity of the magnetic nano-particle of absorption is 10～100 times of single magnetic nano particle daughter nucleus, every absorption one deck just utilizes magnetic separation technique to wash, and forms the magnetic Nano mixture; Said absorption semiconductor fluorescence is nanocrystalline to be, it is nanocrystalline to add semiconductor fluorescence in the magnetic Nano complex solution, utilize the magnetic separation technique washing, adsorb polyelectrolyte again, adsorb the nanocrystalline and polyelectrolyte of semiconductor fluorescence through layer by layer, obtain the fluorescence magnetic nano-complex, the nanocrystalline each adsorptive capacity of semiconductor fluorescence is previous 1～2 times; Said polyelectrolyte multilayer film is polycation electrolyte and the alternate coating of polyanion electrolyte 1～30 layer.
According to the preparation method of the described fluorescence magnetic nano-complex of claim 2, it is characterized in that 3, said magnetic nano-particle is Fe
3O
4Or Fe
2O
3Or CoFe
2O
4Or NiFeO
4Or ZnFe
2O
4Or MnFeO
4Said semiconductor fluorescence is nanocrystalline to be CdS or CdSe or CdTe or ZnS or their doped and compounded thing; Salt in the said absorption polyelectrolyte body is sodium chloride or potassium nitrate.
4, a kind of application of fluorescence magnetic nano-complex of claim 1 is characterized in that, at first adds biological sample in the fluorescence magnetic nanocomposite system, coupling connection under 0～10 ℃ of condition; Carrying out the biological sample multicolor fluorescence afterwards detects or the magnetic resolution of biological sample or the magnetic orientation of biological sample.
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|---|---|---|---|
| CNA031271650A CN1524925A (en) | 2003-09-18 | 2003-09-18 | Fluorescent magnetic nanometer compound, preparing process and application thereof |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1903337A1 (en) | 2006-09-20 | 2008-03-26 | Biocartis SA | Coating for microcarriers |
| CN101952725A (en) * | 2007-12-03 | 2011-01-19 | 国立大学法人东京工业大学 | Biosensing method using coated magnetic microparticles and biosensing device to be used in the method |
| CN104541152A (en) * | 2012-06-15 | 2015-04-22 | 卢米尼克斯股份有限公司 | Apparatus, system, and method for image normalization using a gaussian residual of fit selection criteria |
| CN104840977A (en) * | 2015-03-23 | 2015-08-19 | 济南大学 | Method for preparing magnetic fluorescence composite nano drug carrier |
| CN104844839A (en) * | 2015-03-23 | 2015-08-19 | 济南大学 | Preparation method of magnetic fluorescent composite nanoparticle |
| CN104946256A (en) * | 2015-05-28 | 2015-09-30 | 合肥工业大学 | Rapid preparation method of inorganic compound coated aqueous phase II-VI group quantum dot composite material |
-
2003
- 2003-09-18 CN CNA031271650A patent/CN1524925A/en active Pending
Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1903337A1 (en) | 2006-09-20 | 2008-03-26 | Biocartis SA | Coating for microcarriers |
| WO2008034275A1 (en) * | 2006-09-20 | 2008-03-27 | Biocartis Sa | Coating for microcarriers |
| CN101952725A (en) * | 2007-12-03 | 2011-01-19 | 国立大学法人东京工业大学 | Biosensing method using coated magnetic microparticles and biosensing device to be used in the method |
| CN101952725B (en) * | 2007-12-03 | 2014-12-10 | 多摩川精机株式会社 | Biosensing method using coated magnetic microparticles and biosensing device to be used in the method |
| CN104541152B (en) * | 2012-06-15 | 2017-08-25 | 卢米尼克斯股份有限公司 | Use the image normalization devices, systems, and methods of the Gauss residual error of fitting selection standard |
| CN104541152A (en) * | 2012-06-15 | 2015-04-22 | 卢米尼克斯股份有限公司 | Apparatus, system, and method for image normalization using a gaussian residual of fit selection criteria |
| US9984279B2 (en) | 2012-06-15 | 2018-05-29 | Luminex Corporation | Apparatus, system, and method for image normalization using a gaussian residual of fit selection criteria |
| US10733418B2 (en) | 2012-06-15 | 2020-08-04 | Luminex Corporation | Apparatus, system, and method for image normalization using a Gaussian residual of fit selection criteria |
| CN104840977A (en) * | 2015-03-23 | 2015-08-19 | 济南大学 | Method for preparing magnetic fluorescence composite nano drug carrier |
| CN104844839A (en) * | 2015-03-23 | 2015-08-19 | 济南大学 | Preparation method of magnetic fluorescent composite nanoparticle |
| CN104840977B (en) * | 2015-03-23 | 2017-12-12 | 济南大学 | A kind of preparation method of magnetic fluorescence composite Nano pharmaceutical carrier |
| CN104946256A (en) * | 2015-05-28 | 2015-09-30 | 合肥工业大学 | Rapid preparation method of inorganic compound coated aqueous phase II-VI group quantum dot composite material |
| CN104946256B (en) * | 2015-05-28 | 2016-08-17 | 合肥工业大学 | A kind of inorganic matter is coated with the fast preparation method of aqueous phase II VI race quantum dot composite material |
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