CN105181662A - Qualitative detection method for distribution of functional nanometer particles carried in polysaccharide microspheres - Google Patents

Qualitative detection method for distribution of functional nanometer particles carried in polysaccharide microspheres Download PDF

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CN105181662A
CN105181662A CN201510540038.2A CN201510540038A CN105181662A CN 105181662 A CN105181662 A CN 105181662A CN 201510540038 A CN201510540038 A CN 201510540038A CN 105181662 A CN105181662 A CN 105181662A
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distribution
fitc
nano particle
particles
solution
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CN105181662B (en
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薛伟明
马何平
樊海明
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Northwest University
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Abstract

The invention discloses a qualitative detection method for distribution of functional nanometer particles carried in polysaccharide microspheres. The method more specifically relates to functional nanometer particles which are labeled by fluorescein isothiocyanate (FITC) and are coated by polysaccharide microspheres; at the wavelength of 488 nm, a fluorescence microscope and a laser scanning confocal microscope excite fluorescein to emit green light, so that the distribution of nanometer particles in the polysaccharide microspheres can be determined. Cystamine with primary amino groups at two ends is used for modification of nanometer particles with carboxyl at the surface, thereby providing reactive groups for reaction with FITC, because reaction can be carried out between FITC and specific amino function groups, a labeling of functional nanometer particles with FITC is realized. The invention has the advantages of simple preparation process, mild reaction condition, good stability, preservation convenience and good biocompatibility; according to the invention, visual detection of distribution of nanometer particles in drug carrier is realized, and the method can be widely applied in the field of biological application of nano-material.

Description

The method of a kind of qualitative detection load function nano distribution of particles in polysaccharide microsphere
Technical field
The invention belongs to nano material biologic applications technical field, relate to the method for a kind of qualitative detection load function nano distribution of particles in polysaccharide microsphere.
Background technology
Function nano particle refers to have specific function, as Fe 3o 4nano particles etc., have unique magnetic response performance, can make promptly magnetic response, have a wide range of applications in magnetic heat cure tumour, targeted drug delivery and stimulation-response Co ntrolled release etc. additional magnetic signal.In recent years, the development of pharmaceutical carrier, again for the widespread use of nano particle provides wide development space, by pharmaceutical carrier coated magnetic Fe 3o 4nano particle, both can significantly improve the biocompatibility of nano particle, can utilize again the magnetic performance of nano particle, significantly improves the utilization factor of medicine, has important application value.
Material is the key factor determining pharmaceutical carrier characteristic, and generally, drug carrier material all needs to possess good film-forming property, does not react with encapsulation object, good biocompatibility and the characteristic such as degradability is good.Sodium alginate, shitosan are because of advantages such as its aboundresources, preparation are simple, low prices, simultaneously, the natural polysaccharide hydrogel sphere that sodium alginate-chitosan is formed has three-dimensional netted space structure, can loaded with nano particle and antineoplastic in a large number, therefore, the application of pharmaceutical carrier is subject to extensive concern.
At present, the technique preparing hydrogel microsphere mainly contains emulsion process, interfacial polymerization, phase separation method, spray drying process and electrostatic drop generation etc.And electrostatic drop generation is the microfluidic process that acted on by electric field force, make solution formation diameter much smaller than the high-speed jet of jet size and break to form charged drop, drop makes microballoon have excellent spontaneous monodispersity because carrying the generation repulsive interaction of same sex electric charge in the curing process.Microballoon preparation process is gentle, and simple to operate, microspherulite diameter is controlled, and pattern is good, and envelop rate is high, has broad application prospects.
But, magnetic Fe 3o 4after nano-particles encapsulate enters pharmaceutical carrier, its distribution in carrier inside and the impact on pharmaceutical carrier structure, will directly have influence on the magnetic performance of nano particle and the application in pharmaceutical carrier thereof.People attempt various method and mark nano particle, and to observe it in the distribution of pharmaceutical carrier inside, but its process is complicated, and due to FRET (fluorescence resonance energy transfer), easily causes fluorescent quenching, makes it be restricted at biomedical aspect.
Summary of the invention
For solving the problem, the invention provides the method for a kind of qualitative detection load function nano distribution of particles in polysaccharide microsphere.
For achieving the above object, the technical scheme that the present invention takes is:
S1, by the magnetic Fe of marked by fluorescein isothiocyanate 3o 4it is in the sodium alginate aqueous solution of 1-2% that nano particle joins concentration, stirs, and obtains the mixed liquor that concentration is 0.2 ~ 1.0mgFe/mL;
S2, with electrostatic pulse sessile drop method by the mixed liquor of gained instillation concentration be the CaCl of 1-2% 2in aqueous solution, electrostatic field voltage is 350V, and frequency is 400Hz, metallic nozzle diameter 500 μm, and liquid level, apart from dropwising after 2cm, 1h, leaves standstill 30min, with deionized water rinsing three times, removes the Ca of gel ball excess surface 2+ion, crosses 150 object screen cloth screening Microsphere Size, must carry Fe 3o 4-AED-FITC alginate ball;
S3, gained is carried Fe 3o 4after the acetum overlay film 30-40min of-AED-FITC alginate ball shitosan, deionized water rinsing three times, must be coated with Fe 3o 4the AC microballoon (alginate-chitosan microball) of-FITC;
S4, gained is coated with Fe 3o 4under the AC microballoon of-FITC is placed in fluorescent microscope or laser confocal microscope, wavelength is excite under 488nm, observes the distribution of green fluorescence bright spot, is the distribution of function nano particle in polysaccharide microsphere inside.
Wherein, step S1-S4 all completes under lucifuge condition.
Wherein, the magnetic Fe of described marked by fluorescein isothiocyanate 3o 4nano particle is prepared by following steps:
Step one, by water miscible Fe 3o 4nano particle, be dissolved in 2-(N-morpholine) ethyl sulfonic acid (MES) buffer solution, add 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and N-hydroxy-succinamide (NHS) aqueous solution carries out activation 15-30min, the mass ratio (g/g) of EDC and NHS is the mol ratio of 2: 3-2: 5, EDC and carboxyl is 1: 3 ~ 1: 6;
Step 2, join in above-mentioned solution by the PBS aqueous solution being dissolved with the pH=7.4 of cystamine, the mol ratio of carboxyl and cystamine is 1: 3 ~ 1: 5, on shaking table (250r/min), react 2-4h;
After step 3, reaction terminate, by magnetic separation method, nano particle is separated out from solution, spend deionized water particle three times, obtain Fe 3o 4-AED.By the Fe of gained 3o 4-AED is dissolved in deionized water, regulates pH to 8.8, obtain solution A with the NaOH of 0.5mol/L;
Step 4,1mg fluorescein isothiocynate is dissolved in 1mL dimethyl sulfoxide (DMSO), be diluted to after concentration is 50 μ g/mL by the PBS phosphate buffered solution of pH=7.4, mix with solution A, the mol ratio of fluorescein and primary amino radical is 1: 60 ~ 1: 100, under room temperature, shakes bed reaction 4h in 150r/min, after reaction terminates, carry out Magneto separate according to step 3, wash three times with ethanol, obtain Fe 3o 4-AED-FITC.
Wherein, described Fe 3o 4the particle diameter of nano particle is 3 ~ 100nm.
Wherein, the metallic nozzle diameter that impulse electric field sessile drop method adopts is 500 μm.
Wherein, described nano particle is the function nano particle of surface containing carboxyl.
Wherein, described function nano particle is Quantum dots CdS, CdSe, CdTe or ZnSe, Fe 3o 4or MFe 2o 4, wherein, M is metal M n, Zn, Co or Ni.
Wherein, when continuing overlay film with shitosan to gel ball, the volume ratio of gel ball and chitosan solution is 1: 4.
The present invention has following beneficial effect:
Adopt FITC flag F e 3o 4nano particle, by cystamine to Fe 3o 4nano particle carries out finishing, for next step grafting FITC provides reaction amino.Compared with prior art, grafting efficiency of the present invention is high, and reaction product is stablized, and is easy to preserve, not easily fluorescent quenching phenomenon occurs because of resonance energy transfer.The drug carrier material adopted is alginate/shitosan (AC) microballoon, this microballoon has three-dimensional netted space structure, can loaded with nano particle and antineoplastic in a large number, have envelop rate advantages of higher, the development for pharmaceutical carrier provides vast development space.
Accompanying drawing explanation
The water-soluble Fe of 19nm of Figure 1A embodiment of the present invention 1 3o 4the TEM figure of nano particle.
The water-soluble Fe of 19nm of Figure 1B embodiment of the present invention 1 3o 4the dynamic light scattering of nano particle characterizes.
The Fe of Fig. 1 C embodiment of the present invention 1 3o 4nano particle and Fe 3o 4the FTIR figure of-AED.
The Fe of Fig. 2 embodiment of the present invention 2 3o 4-AED and Fe 3o 4the ultraviolet-visible absorption spectroscopy figure of-AED-FITC.
The concentration of Fig. 3 A and the 3B embodiment of the present invention 3 is year Fe of 0.38mgFe/mL 3o 4the microphotograph of AC microballoon of-AED-FITC and the grain size distribution of microballoon.
The concentration of Fig. 3 C embodiment of the present invention 3 is year Fe of 0.38mgFe/mL 3o 4the AC fluorescent microscopy images of-AED-FITC.
The concentration of Fig. 3 D embodiment of the present invention 3 is year Fe of 0.38mgFe/mL 3o 4the laser confocal microscope photo of the AC microballoon of-AED-FITC.
The concentration of Fig. 4 A embodiment of the present invention 4 is year Fe of 0.80mgFe/mL 3o 4the AC microsphere laser Laser Scanning Confocal Microscope photo of-AED-FITC.
The concentration of Fig. 4 B embodiment of the present invention 4 is year Fe of 0.80mgFe/mL 3o 4the z-axis distribution laser confocal microscope photo of the AC microballoon of-AED-FITC.
Fig. 5 is the magnetic Fe that in the embodiment of the present invention, fluorescein isothiocynate (FITC) marks 3o 4nano particle (Fe 3o 4-AED-FITC) preparation flow figure.
Embodiment
In order to make objects and advantages of the present invention clearly understand, below in conjunction with embodiment, the present invention is further elaborated.Should be appreciated that specific embodiment described herein only in order to explain the present invention, be not intended to limit the present invention.
Embodiment 1
Fe is modified with cystamine 3o 4the method of nano particle:
19nmFe 3o 4nano particle prepare list of references: NatureMaterials.2004,3,891-895.
Oil phase nano particle turn water list of references: Xi'an Goldmag Nanobiotech Co., Ltd., reference number of a document GMAG-QI-PD-O2O.
5mL is dissolved with the Fe of 19nm 3o 4the MES buffer solution (pH=6) in 50mL plastic centrifuge tube of nano particle (be 0.03mmol/mL containing carboxyl concentration), add 0.5mLEDC aqueous solution (concentration is 2mmol/mL), on the shaking table of 150r/min, vibration 15min; Add 0.5mLNHS aqueous solution (concentration is 10mmol/mL) in above-mentioned solution; Then the PBS buffer solution (pH=7.4) of 25mL containing cystamine (concentration is 0.03mmol/mL) is added, react on the shaking table of 150r/min, cessation reaction after 2h, pass through magnetic separation method, nano particle is separated out from solution, spend deionized water, repeat aforesaid operations three times, final reaction product (Fe 3o 4-AED) be dissolved in 2mL deionized water.
About 15uL is dispersed with 19nmFe 3o 4it is online that the aqueous solution of nano particle drops in the Cu being coated with carbon film, after natural drying, does transmission electron microscope, as shown in Figure 1A.
Get 1 ~ 1.5mLFe 3o 4the aqueous solution of nano particle, uses MalvernNanoZetaSizerAnalyzer to carry out light scattering test hydrated radius, known 19nmFe from Figure 1B 3o 4nanoparticulate dispersed average hydrated radius after aqueous solution is 113.8nm.
By 2mLFe 3o 4the aqueous solution of nano particle and be grafted with the Fe of cystamine 3o 4nano particle (Fe 3o 4-AED) aqueous solution in vacuum drying chamber at 30 DEG C, vacuum drying 12h, after becoming pressed powder, characterize with Fourier transform infrared spectroscopy (FTIR), result as shown in Figure 1 C, can see Fe 3o 4-COOH nano particle is 1588cm in wave number -1there is-COOH peak in position, is 593cm in wave number -1position there is Fe-O peak; Fe after grafting cystamine 3o 4-AED nano particle is 1632cm in wave number -1there is-CO-NH-peak in position, is 3272cm in wave number -1position appearance-NH 2peak, illustrates the successful grafting of cystamine, and has free amino, is beneficial to next step and FITC reacts.
Embodiment 2
Be marked with the Fe of FITC 3o 4the preparation of-AED-FITC:
Be dissolved in by 1mg fluorescein isothiocynate (FITC) in 1mL dimethyl sulfoxide (DMSO) (DMSO), being diluted to concentration by the PBS phosphate buffered solution of pH=7.4 is 50 μ g/mL; This solution is joined containing Fe 3o 4(containing amino 0.17mmol, regulate pH to 8.8 with the NaOH of 0.5mol/L) in the 20mL aqueous solution of-AED, the mol ratio of fluorescein and primary amino radical is 1: 60 ~ 1: 100, under room temperature, on the shaking table of 150r/min, carry out reaction 4h.After reaction terminates, by magnetic separation method, nano particle is separated out from solution, spend ethanol washing granule, repeat this operation three times, final product Fe 3o 4-AED-FITC deposits in PBS buffer solution, keeps in Dark Place at 4 DEG C.
Reaction product Fe 3o 4the mensuration of-AED-FITC concentration: pipette 200 μ L above-mentioned containing Fe 3o 4the PBS buffer solution of-FITC, add 1mL hypochlorous acid, 3mL nitric acid, nitre solution 3 ~ 4h is carried out at 130 DEG C, after leaving standstill a night, proceed in 10mL volumetric flask, add deionized water constant volume, the content of ferric ion in this aqueous solution is surveyed with inductively coupled plasma (InductivelyCoupledPlasma, ICP).
Get 1 ~ 1.5mLFe 3o 4-AED and Fe 3o 4the aqueous solution of-AED-FITC, ultraviolet-visible absorption spectroscopy (UV-VisAbsorptionspectrum) is used to carry out the peak position test of FITC, as shown in Figure 2, by the contrast of two curves, there is the absorption peak of FITC in the position that wave number is 503nm in the nano particle after can obviously finding out grafting FITC, demonstrates the successful grafting of FITC; It is the characteristic absorption peak of cystine linkage in the peak position cystamine of 224nm position appearance in wave number.
Embodiment 3
Carry the preparation that iron is the AC microballoon of 0.38mgFe/mL:
Fe is prepared by the method for embodiment 1 3o 4-AED-FITC nano particle aqueous solution.
Be the Fe of 3mg by iron-holder 3o 4-AED-FITC is dissolved in 0.5mL deionized water, mixes with sodium alginate aqueous solution, after mixing, and Fe 3o 4the concentration of-FITC is 0.6mgFe/mL, and the concentration of sodium alginate is 1.2%, w/v; By electrostatic field pulse sessile drop method, sodium alginate aqueous solution is instilled CaCl 2in aqueous solution (concentration is 2%, w/v), form hydrogel microsphere, electrostatic field voltage is 350V, and frequency is 400Hz, metallic nozzle diameter 500 μm, and liquid level is apart from 2cm; Dropwise after 1h, leave standstill 30min, remove unnecessary Ca three times with deionized water rinsing 2+ion, with 150 object screen cloth screening Microsphere Size; Acetic acid (2%, the v/v) solution adding shitosan (0.5%, w/v) carries out overlay film, and the volume ratio of gel ball and chitosan solution is 1: 4,30min, with deionized water rinsing three times, must be coated with Fe 3o 4the AC microballoon of-FITC.
The preparation of broken capsule liquid: the NaHCO that basic composition is 0.2mol/L of broken capsule liquid 3with the sodium citrate (Na of 0.06mol/L 3c 6h 5o 712H 2o) aqueous solution, pH=7.8 ~ 8.2.
Get 0.5mLAC microballoon, will break capsule liquid and AC microballoon and mix with volume ratio 10: 1, manual jolting, micro-capsule all dissolves in 30s.Add 1mL hypochlorous acid and 3mL nitric acid in solution after broken capsule, at 130 DEG C, carry out nitre solution 7 ~ 8h, with the content of Fe in ICP test solution.By calculating, this microballoon has higher envelop rate, reaches 59.7%.
Get the aqueous solution of 1 ~ 1.5mLAC microballoon, use fluorescent microscope and OlympusFluoviewFV1000 laser confocal microscope to carry out Fluorescent Characterization.
Fig. 3 A and 3B is for carrying Fe 3o 4the AC microballoon photo of-AED-FITC and concentration are year Fe of 0.38mgFe/mL 3o 4the AC microballoon simple microscope photo of-AED-FITC and grain size distribution.As can be seen from the figure, the particle size distribution range of this microballoon is narrower, and mean grain size is 428 μm.Fig. 3 C and 3D is concentration is that 0.38mgFe/mL carries Fe 3o 4the fluorescent microscopy images of the AC microballoon of-FITC and laser confocal microscope photo, as can be seen from two figure, the position of green light is the Fe of FITC mark 3o 4the position of nano particle, Fe 3o 4the center and peripheral of nano particle all has distribution and distribution is more even.
Embodiment 4
Carry the preparation that iron is the AC microballoon of 0.80mgFe/mL:
Be the Fe of 6.5mg by iron-holder 3o 4-FITC is dissolved in 0.5mL deionized water, mixes, Fe with sodium alginate aqueous solution (concentration is 1.2%, w/v) 3o 4the concentration of-FITC is 1.3mgFe/mL; By electrostatic field pulse sessile drop method, sodium alginate aqueous solution is instilled CaCl 2in aqueous solution (concentration is 2%, w/v), form hydrogel microsphere, electrostatic field voltage is 300V, and frequency is 400Hz, metallic nozzle diameter 500 μm, and liquid level is apart from 2cm; Dropwise after 1h, leave standstill 30min, remove unnecessary Ca three times with deionized water rinsing 2+ion, with 150 object screen cloth screening Microsphere Size; Acetic acid (2%, the v/v) solution adding shitosan (0.5%, w/v) carries out overlay film, and the volume ratio of gel ball and chitosan solution is 1: 4,30min, with deionized water rinsing three times, must be coated with Fe 3o 4the AC microballoon of-FITC.
Broken capsule liquid in use-case 3 carries out brokenly capsule, tests the concentration of Fe with ICP, and by calculating, the envelop rate of this microballoon is 50.0%.
Get the aqueous solution of 1 ~ 1.5mLAC microballoon, use fluorescent microscope and OlympusFluoviewFV1000 laser confocal microscope to carry out Fluorescent Characterization.
Fig. 4 A to be concentration be 0.80mgFe/mL's carries Fe 3o 4the AC microsphere laser Laser Scanning Confocal Microscope photo of-AED-FITC, compares with Fig. 3 D, and the green glow density ratio in this microballoon is comparatively large, and Fe in this microballoon is described 3o 4the concentration of nano particle is larger.
Fig. 4 B to be concentration be 0.80mgFe/mL's carries Fe 3o 4the z-axis distribution laser confocal microscope photo of the AC microballoon of-AED-FITC, as can be seen from the figure from the top-to-bottom of this microballoon, green glow density is ascending to little, and this figure can see Fe clearly 3o 4nano particle is in the distribution of microballoon inside.
The above is only the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, under the premise without departing from the principles of the invention; can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.

Claims (8)

1. a method for qualitative detection load function nano distribution of particles in polysaccharide microsphere, is characterized in that, comprise the following steps:
S1, by the magnetic Fe of marked by fluorescein isothiocyanate 3o 4it is in the sodium alginate aqueous solution of 1-2% that nano particle joins concentration, stirs, and obtains the mixed liquor that concentration is 0.2 ~ 1.0mgFe/mL;
S2, with electrostatic pulse sessile drop method by the mixed liquor of gained instillation concentration be the CaCl of 1-2% 2in aqueous solution, electrostatic field voltage is 350V, and frequency is 400Hz, metallic nozzle diameter 500 μm, and liquid level, apart from dropwising after 2cm, 1h, leaves standstill 30min, with deionized water rinsing three times, removes the Ca of gel ball excess surface 2+ion, crosses 150 object screen cloth screening Microsphere Size, must carry Fe 3o 4-AED-FITC alginate ball;
S3, gained is carried Fe 3o 4after the acetum overlay film 30-40min of-AED-FITC alginate ball shitosan, deionized water rinsing three times, must be coated with Fe 3o 4the AC microballoon of-FITC;
S4, gained is coated with Fe 3o 4under the AC microballoon of-FITC is placed in fluorescent microscope or laser confocal microscope, wavelength is excite under 488nm, observes the distribution of green fluorescence bright spot, is the distribution of function nano particle in polysaccharide microsphere inside.
2. the method for a kind of qualitative detection load according to claim 1 function nano distribution of particles in polysaccharide microsphere, it is characterized in that, step S1-S4 all completes under lucifuge condition.
3. the method for a kind of qualitative detection load according to claim 1 function nano distribution of particles in polysaccharide microsphere, is characterized in that, the magnetic Fe of described marked by fluorescein isothiocyanate 3o 4nano particle is prepared by following steps:
Step one, by water miscible Fe 3o 4nano particle, be dissolved in 2-(N-morpholine) ethyl sulfonic acid (MES) buffer solution, add 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride and N-hydroxy-succinamide aqueous solution carries out activation 15-30min, the mass ratio of EDC and NHS is the mol ratio of 2: 3-2: 5, EDC and carboxyl is 1: 3 ~ 1: 6;
Step 2, join in above-mentioned solution by the PBS aqueous solution being dissolved with the pH=7.4 of cystamine, the mol ratio of carboxyl and cystamine is 1: 3 ~ 1: 5, and in shaking table, 250r/min reacts 2-4h;
After step 3, reaction terminate, by magnetic separation method, nano particle is separated out from solution, spend deionized water particle three times, obtain Fe 3o 4-AED, by the Fe of gained 3o 4-AED is dissolved in deionized water, regulates pH to 8.8, obtain solution A with the NaCH of 0.5mol/L;
Step 4,1mg fluorescein isothiocynate is dissolved in 1mL dimethyl sulfoxide (DMSO), be diluted to after concentration is 50 μ g/mL by the PBS phosphate buffered solution of pH=7.4, mix with solution A, the mol ratio of fluorescein and primary amino radical is 1: 60 ~ 1: 100, under room temperature, shakes bed reaction 4h in 150r/min, after reaction terminates, carry out Magneto separate according to step 3, wash three times with ethanol, obtain Fe 3o 4-AED-FITC.
4. the method for a kind of qualitative detection load according to claim 3 function nano distribution of particles in polysaccharide microsphere, is characterized in that, described Fe 3o 4the particle diameter of nano particle is 3 ~ 100nm.
5. the method for a kind of qualitative detection load according to claim 3 function nano distribution of particles in polysaccharide microsphere, it is characterized in that, the metallic nozzle diameter that impulse electric field sessile drop method adopts is 500 μm.
6. the method for a kind of qualitative detection load according to claim 3 function nano distribution of particles in polysaccharide microsphere, is characterized in that, described nano particle is the function nano particle of surface containing carboxyl.
7. the method for a kind of qualitative detection load according to claim 6 function nano distribution of particles in polysaccharide microsphere, it is characterized in that, described function nano particle is Quantum dots CdS, CdSe, CdTe or ZnSe, Fe 3o 4or MFe 2o 4, wherein, M is metal M n, Zn, Co or Ni.
8. the method for a kind of qualitative detection load according to claim 3 function nano distribution of particles in polysaccharide microsphere, is characterized in that, when continuing overlay film with shitosan to gel ball, the volume ratio of gel ball and chitosan solution is 1: 4.
CN201510540038.2A 2015-08-24 2015-08-24 A kind of method that qualitative detection is supported on function nano distribution of particles in polysaccharide microsphere Expired - Fee Related CN105181662B (en)

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115041143A (en) * 2022-04-02 2022-09-13 中国医学科学院基础医学研究所 Magnetic polymer, preparation method, kit and application
CN115058055A (en) * 2022-04-02 2022-09-16 中国医学科学院基础医学研究所 Cystamine modified functionalized magnetic material and preparation method and application thereof
CN115812700A (en) * 2021-12-31 2023-03-21 中山大学 Nano pesticide and preparation method thereof

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4267235A (en) * 1979-03-19 1981-05-12 California Institute Of Technology Polyglutaraldehyde microspheres
CN1560633A (en) * 2004-02-19 2005-01-05 上海交通大学 Preparation method of quantum point microspheric for bio-medical fluorescence probe
US20060293396A1 (en) * 2005-01-14 2006-12-28 Eastman Kodak Company Amine polymer-modified nanoparticulate carriers
CN102964608A (en) * 2012-11-15 2013-03-13 中北大学 Preparation method of carbon quantum dot containing calcium alginate gel for detecting copper ions
CN103333675A (en) * 2013-03-18 2013-10-02 杭州德安奇生物工程有限公司 Fluorescent microsphere suitable for rapid immunology detection and preparation method thereof
CN103525405A (en) * 2013-10-21 2014-01-22 北京理工大学 Magnetic fluorescent difunctional nano material based on natural polymer and preparation method thereof

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4267235A (en) * 1979-03-19 1981-05-12 California Institute Of Technology Polyglutaraldehyde microspheres
CN1560633A (en) * 2004-02-19 2005-01-05 上海交通大学 Preparation method of quantum point microspheric for bio-medical fluorescence probe
US20060293396A1 (en) * 2005-01-14 2006-12-28 Eastman Kodak Company Amine polymer-modified nanoparticulate carriers
CN102964608A (en) * 2012-11-15 2013-03-13 中北大学 Preparation method of carbon quantum dot containing calcium alginate gel for detecting copper ions
CN103333675A (en) * 2013-03-18 2013-10-02 杭州德安奇生物工程有限公司 Fluorescent microsphere suitable for rapid immunology detection and preparation method thereof
CN103525405A (en) * 2013-10-21 2014-01-22 北京理工大学 Magnetic fluorescent difunctional nano material based on natural polymer and preparation method thereof

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
NING SUI等: "Magnetic and optical properties of Ag@SiO2-FITC-Fe3O4 hybrid nanoparticles", 《MATERIALS SCIENCE AND ENGINEERING B》 *
刘萍等: "脉冲电场制备磁性Fe3O4海藻酸盐微球的工艺研究", 《食品与发酵工业》 *
范妮等: "壳聚糖-海藻酸钙微球荧光标记反应对微球膜强度影响的研究", 《四川师范大学学报》 *
范妮等: "壳聚糖荧光标记反应研究", 《云南大学学报》 *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115812700A (en) * 2021-12-31 2023-03-21 中山大学 Nano pesticide and preparation method thereof
CN115041143A (en) * 2022-04-02 2022-09-13 中国医学科学院基础医学研究所 Magnetic polymer, preparation method, kit and application
CN115058055A (en) * 2022-04-02 2022-09-16 中国医学科学院基础医学研究所 Cystamine modified functionalized magnetic material and preparation method and application thereof
CN115058055B (en) * 2022-04-02 2023-10-20 中国医学科学院基础医学研究所 Cystamine modified functional magnetic material and preparation method and application thereof

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