CN102618257B - A surface coating method for upconversion fluorescent nanomaterials - Google Patents
A surface coating method for upconversion fluorescent nanomaterials Download PDFInfo
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- CN102618257B CN102618257B CN201210060849.9A CN201210060849A CN102618257B CN 102618257 B CN102618257 B CN 102618257B CN 201210060849 A CN201210060849 A CN 201210060849A CN 102618257 B CN102618257 B CN 102618257B
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- 238000006243 chemical reaction Methods 0.000 claims abstract description 50
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims abstract description 46
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- 238000000034 method Methods 0.000 claims abstract description 22
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Abstract
本发明属于光子学及材料学交叉技术领域，基于研究简单高效、生物兼容性好的上转换荧光纳米材料包覆方法的思路，采用牛血清蛋白作为表面活性剂，在超声分散的四氢呋喃和水混合液中，利用反溶剂法对上转换荧光纳米材料表面进行牛血清蛋白包覆，随后加入戊二醛将纳米材料表面的牛血清蛋白分子进行联接，提高表面包覆层的稳定性。其中用到的表面包覆剂是生物体中提取的蛋白分子，具有极高的生物兼容性，对于利用上转换荧光纳米材料进行活体成像研究的开展创造了便利。
The invention belongs to the interdisciplinary technical field of photonics and materials science. Based on the idea of researching simple, efficient and biocompatible up-conversion fluorescent nanomaterial coating methods, bovine serum albumin is used as a surfactant and mixed in ultrasonically dispersed tetrahydrofuran and water. In the solution, the surface of the upconversion fluorescent nanomaterial was coated with bovine serum albumin by anti-solvent method, and then glutaraldehyde was added to link the bovine serum albumin molecules on the surface of the nanomaterial to improve the stability of the surface coating layer. The surface coating agent used here is a protein molecule extracted from a living body, which has extremely high biocompatibility and creates convenience for the development of in vivo imaging research using up-conversion fluorescent nanomaterials.
Description
Technical field
The invention belongs to photonics and materialogy interleaving techniques field, be specifically related to the surface coating method of up-conversion fluorescence nano material.
Background technology
Up-conversion fluorescence nano material is as the novel fluorescent agent of a class, has that near infrared excites, high signal to noise ratio, the advantages such as hypotoxicity, bio-imaging particularly living body biological imaging side mask have unique advantage.The synthetic method of existing up-conversion fluorescence nano material is mainly to adopt oleic acid as tensio-active agent, and the up-conversion fluorescence nano material of preparing can be dispersed in organic solvent, is but difficult for being dispersed in water surrounding.Therefore, by up-conversion fluorescence Application of micron in bio-imaging, just need more be coated it, improve on the one hand the stability of material in the aqueous solution, improve on the one hand its biological suitability.At present common up-conversion fluorescence nano material is coated is mainly the coated two kinds of methods of coated with silica and high molecular polymer.The ultimate principle of coated with silica is to utilize tetraethoxy, at nano-material surface, self hydrolysis reaction formation layer of silicon dioxide occurs, and main chemical reactions equation is as follows:
Si（OC
2H
5）+2H
2O
SiO
2+4 C
2H
5OH
Up-conversion fluorescence nano material after coated with silica can be good be dispersed in the aqueous solution, on silicon dioxide layer surface, can carry out easily the modification of functionalization group, the application coated at material surfaces such as organic dye, metallic particles, quantum dots is also very extensive simultaneously.High molecular polymer is coated is mainly to adopt coprecipitation method, at the coated hydrophilic high molecular polymer of one deck of nano-material surface, thereby improves stability and the bio-compatibility of nano material in the aqueous solution.It is pointed out that above-mentioned two kinds of method for coating all exist some shortcomings:
1. the existence of silicon dioxide layer has increased the size of material, is unfavorable for material circulation in vivo, easily by immunity system, caught, and also comparatively complicated in the program of nano grain surface coated silica, there is suitable difficulty and diversity in reaction process.
2. adopt coprecipitation method effectively not connect the coated high molecular polymer of nano-material surface is intermolecular, the less stable of surface coating layer, is difficult to adapt to complicated organism environment.
3. no matter being silicon-dioxide or high molecular polymer, is not the material having in organism self, has certain limitation aspect bio-compatibility.
Based on above some consideration, study a kind of more simple up-conversion fluorescence nano material method for coating efficient, that bio-compatibility is high and seem very necessary.
Summary of the invention
The present invention is the thinking of the up-conversion fluorescence nano material method for coating based on research is simple efficiently, bio-compatibility is good just, adopt bovine serum albumin as tensio-active agent, in the tetrahydrofuran (THF) and water mixed liquid of ultrasonic dispersion, utilizing anti-solvent method to carry out bovine serum albumin to up-conversion fluorescence nano-material surface is coated, add subsequently glutaraldehyde that the bovine serum albumin molecule of nano-material surface is connected, improve the stability of surface coating layer.The surperficial coating of wherein using is the protein molecular extracting in organism, has high bio-compatibility, for utilizing up-conversion fluorescence nano material to carry out carrying out of living imaging research, has created facility.
Up-conversion fluorescence nano material method for coating step of the present invention is as follows:
1. according to quality ratio up-conversion fluorescence nano material (UCNPs) powder: bovine serum albumin (BSA) powder is that 1:1～1.1 take these two kinds of powder and put into respectively two centrifuge tubes.In the centrifuge tube that UCNPs powder is housed, add a certain amount of tetrahydrofuran (THF) ultrasonic dispersion 1～2 minute, obtain the tetrahydrofuran solution that UCNPs concentration is 1～1.1mg/mL.In the centrifuge tube that BSA powder is housed, add a certain amount of deionized water ultrasonic dispersion 1～2 minute, obtain the aqueous solution that BSA concentration is 1～1.1mg/mL.
2. the centrifuge tube that UCNPs solution is housed is placed in to ultrasonic cleaning machine, the aqueous solution of BSA is dropwise added wherein.In this process, BSA molecule, by the anti-solvation in tetrahydrofuran (THF)-water mixed liquid, is coated to up-conversion fluorescence nano material outside surface.
3. after the BSA aqueous solution dropwises, in mixing solutions, add 8～10 microlitre glutaraldehyde solutions, then, by solution left standstill 24～36 hours, make glutaraldehyde follow the full cross-linked reaction of BSA molecule, form and stablize fine and close coating layer.
4. pair mixing solutions carries out centrifugal treating, gets bottom settlings and is re-dispersed in deionized water, and repeated centrifugation, dispersion process, be then placed in air drying by the precipitation finally obtaining, and can obtain the coated up-conversion fluorescence nano material pressed powder of BSA.
Realizing the coated required reagent of up-conversion fluorescence nano material method of above-mentioned BSA comprises: oil soluble up-conversion fluorescence nano material powder, bovine serum albumin powder, glutaraldehyde, tetrahydrofuran (THF).Required experimental installation equipment comprises: ultrasonic cleaner, whizzer, liquid-transfering gun.
The bovine serum albumin that the inventive method adopts, as the outer coating of up-conversion fluorescence nano material, utilizes glutaraldehyde that bovine serum albumin is molecule crosslinked, forms fine and close outer clad structure, makes nano material have stable chemical property.Meanwhile, bovine serum albumin, as naturally occurring protein molecular in a kind of organism, has better bio-compatibility than traditional coating materials such as silicon-dioxide or polyoxyethylene glycol (PEG), in bio-imaging field, has broad application prospects.
Accompanying drawing explanation
Fig. 1 is principle and the schema of the inventive method.
Fig. 2 is the fluorescence emission spectrum of the up-conversion fluorescence nano material aqueous solution that is coated with bovine serum albumin prepared of the inventive method under 980 nm laser excitations.Illustration is the aqueous solution (massfraction is 1mg/mL) of the up-conversion fluorescence nano material that is coated with bovine serum albumin prepared of the inventive method digital camera photo under white light and 980nm laser radiation.
Embodiment 1
Take respectively 10 milligrams of up-conversion fluorescence nano materials (UCNPs) and 10 milligrams of bovine serum albumins (BSA) powder packs in two 50mL centrifuge tubes, to being equipped with in the centrifuge tube of UCNPs, add 10 milliliters of tetrahydrofuran solutions, to being equipped with in the centrifuge tube of BSA, add 10 ml deionized water, then two centrifuge tubes are put in ultrasonic cleaning machine and take out after ultrasonic 1 minute.The centrifuge tube that UCNPs solution is housed is reentered in ultrasonic cleaning machine, with liquid-transfering gun, BSA solution is dropwise joined in the centrifuge tube that UCNPs solution is housed.After BSA solution dropwises, in mixing solutions, add 8 microlitre glutaraldehyde solutions, then by solution left standstill 24 hours.Question response is isolated throw out by centrifuging completely, with deionized water, repeatedly cleans, and is placed in after air drying, can be coated with the up-conversion fluorescence nano material pressed powder of bovine serum albumin.
Embodiment 2
Take respectively 10 milligrams of up-conversion fluorescence nano materials (UCNPs) and 11 milligrams of bovine serum albumins (BSA) powder packs in two 50mL centrifuge tubes, to being equipped with in the centrifuge tube of UCNPs, add 10 milliliters of tetrahydrofuran solutions, to being equipped with in the centrifuge tube of BSA, add 11 ml deionized water, then two centrifuge tubes are put in ultrasonic cleaning machine and take out after ultrasonic 2 minutes.The centrifuge tube that UCNPs solution is housed is reentered in ultrasonic cleaning machine, with liquid-transfering gun, BSA solution is dropwise joined in the centrifuge tube that UCNPs solution is housed.After BSA solution dropwises, in mixing solutions, add 10 microlitre glutaraldehyde solutions, then by solution left standstill 36 hours.Question response is isolated throw out by centrifuging completely, with deionized water, repeatedly cleans, and is placed in after air drying, can be coated with the up-conversion fluorescence nano material pressed powder of bovine serum albumin.
Embodiment 3
Take respectively 10 milligrams of up-conversion fluorescence nano materials (UCNPs) and 10.5 milligrams of bovine serum albumins (BSA) powder packs in two 50mL centrifuge tubes, to being equipped with in the centrifuge tube of UCNPs, add 10 milliliters of tetrahydrofuran solutions, to being equipped with in the centrifuge tube of BSA, add 10.5 ml deionized water, then two centrifuge tubes are put in ultrasonic cleaning machine and take out after ultrasonic 1.5 minutes.The centrifuge tube that UCNPs solution is housed is reentered in ultrasonic cleaning machine, with liquid-transfering gun, BSA solution is dropwise joined in the centrifuge tube that UCNPs solution is housed.After BSA solution dropwises, in mixing solutions, add 9 microlitre glutaraldehyde solutions, then by solution left standstill 30 hours.Question response is isolated throw out by centrifuging completely, with deionized water, repeatedly cleans, and is placed in after air drying, can be coated with the up-conversion fluorescence nano material pressed powder of bovine serum albumin.
Embodiment 4
Take respectively 10 milligrams of up-conversion fluorescence nano materials (UCNPs) and 10 milligrams of bovine serum albumins (BSA) powder packs in two 50mL centrifuge tubes, to being equipped with in the centrifuge tube of UCNPs, add 11 milliliters of tetrahydrofuran solutions, to being equipped with in the centrifuge tube of BSA, add 11 ml deionized water, then two centrifuge tubes are put in ultrasonic cleaning machine and take out after ultrasonic 1 minute.The centrifuge tube that UCNPs solution is housed is reentered in ultrasonic cleaning machine, with liquid-transfering gun, BSA solution is dropwise joined in the centrifuge tube that UCNPs solution is housed.After BSA solution dropwises, in mixing solutions, add 8 microlitre glutaraldehyde solutions, then by solution left standstill 24 hours.Question response is isolated throw out by centrifuging completely, with deionized water, repeatedly cleans, and is placed in after air drying, can be coated with the up-conversion fluorescence nano material pressed powder of bovine serum albumin.
Embodiment 5
Take respectively 10.5 milligrams of up-conversion fluorescence nano materials (UCNPs) and 10.5 milligrams of bovine serum albumins (BSA) powder packs in two 50mL centrifuge tubes, to being equipped with in the centrifuge tube of UCNPs, add 10 milliliters of tetrahydrofuran solutions, to being equipped with in the centrifuge tube of BSA, add 10 ml deionized water, then two centrifuge tubes are put in ultrasonic cleaning machine and take out after ultrasonic 1.5 minutes.The centrifuge tube that UCNPs solution is housed is reentered in ultrasonic cleaning machine, with liquid-transfering gun, BSA solution is dropwise joined in the centrifuge tube that UCNPs solution is housed.After BSA solution dropwises, in mixing solutions, add 10 microlitre glutaraldehyde solutions, then by solution left standstill 24 hours.Question response is isolated throw out by centrifuging completely, with deionized water, repeatedly cleans, and is placed in after air drying, can be coated with the up-conversion fluorescence nano material pressed powder of bovine serum albumin.
Embodiment 6
Take respectively 10 milligrams of up-conversion fluorescence nano materials (UCNPs) and 10 milligrams of bovine serum albumins (BSA) powder packs in two 50mL centrifuge tubes, to being equipped with in the centrifuge tube of UCNPs, add 10 milliliters of tetrahydrofuran solutions, to being equipped with in the centrifuge tube of BSA, add 10 ml deionized water, then two centrifuge tubes are put in ultrasonic cleaning machine and take out after ultrasonic 2 minutes.The centrifuge tube that UCNPs solution is housed is reentered in ultrasonic cleaning machine, with liquid-transfering gun, BSA solution is dropwise joined in the centrifuge tube that UCNPs solution is housed.After BSA solution dropwises, in mixing solutions, add 9 microlitre glutaraldehyde solutions, then by solution left standstill 36 hours.Question response is isolated throw out by centrifuging completely, with deionized water, repeatedly cleans, and is placed in after air drying, can be coated with the up-conversion fluorescence nano material pressed powder of bovine serum albumin.
Below content of the present invention is described further.
As shown in Figure 1, BSA molecule by the anti-solvation in tetrahydrofuran (THF)-water mixed liquid, was coated to up-conversion fluorescence nano material outside surface before this, then passed through the crosslinked action of glutaraldehyde molecule, formed and stablized fine and close coating layer.
As shown in Figure 2, the up-conversion fluorescence nano material that is coated with bovine serum albumin of preparing based on the inventive method has good up-conversion fluorescence effect under 980nm laser apparatus excites.And show that the up-conversion fluorescence nano material that is coated with bovine serum albumin based on the inventive method has good dispersiveness in the aqueous solution.
Claims (1)
1. a surface coating method for up-conversion fluorescence nano material, is characterized in that the method has adopted the surperficial coating of bovine serum albumin as material, and comprises the steps:
Step 1. is according to quality ratio up-conversion fluorescence nano material powder: bovine serum albumin powder is that 1:1～1.1 take these two kinds of powder and put into respectively two centrifuge tubes, in the centrifuge tube that up-conversion fluorescence nano material powder is housed, add a certain amount of tetrahydrofuran (THF) ultrasonic dispersion 1～2 minute, obtain the tetrahydrofuran solution that up-conversion fluorescence nano material concentration is 1～1.1mg/mL; In the centrifuge tube that bovine serum albumin powder is housed, add a certain amount of deionized water ultrasonic dispersion 1～2 minute, obtain the aqueous solution that bovine serum albumin concentration is 1～1.1mg/mL;
Step 2. is placed in ultrasonic cleaning machine by the centrifuge tube that up-conversion fluorescence nano material solution is housed, and the aqueous solution of bovine serum albumin is dropwise added wherein; In this process, bovine serum albumin molecule, by the anti-solvation in tetrahydrofuran (THF)-water mixed liquid, is coated to up-conversion fluorescence nano material outside surface;
Step 3., after the bovine serum albumin aqueous solution dropwises, adds 8～10 microlitre glutaraldehyde solutions in mixing solutions, then, by solution left standstill 24～36 hours, makes glutaraldehyde follow the full cross-linked reaction of bovine serum albumin molecule, forms and stablizes fine and close coating layer;
Step 4. pair mixing solutions carries out centrifugal treating, getting bottom settlings is re-dispersed in deionized water, repeated centrifugation, dispersion process, be then placed in air drying by the precipitation finally obtaining, and can obtain the coated up-conversion fluorescence nano material pressed powder of bovine serum albumin.
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| CN1521231A (en) * | 2003-01-23 | 2004-08-18 | 中国人民解放军军事医学科学院微生物 | Irradiant material with surface modification and activation |
| US7524630B2 (en) * | 2002-04-22 | 2009-04-28 | University Of Florida Research Foundation, Inc. | Functionalized nanoparticles and methods of use |
| CN102295932A (en) * | 2011-07-05 | 2011-12-28 | 武汉大学 | Method for preparing ZnSe quantum dot marked bovine serum albumin fluorescent probe |
| CN102366632A (en) * | 2011-08-22 | 2012-03-07 | 长春工业大学 | Paramagnetic metal complex functionalized fluorogold nano-cluster magnetic resonance and fluorescence imaging contrast agent |
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| US7524630B2 (en) * | 2002-04-22 | 2009-04-28 | University Of Florida Research Foundation, Inc. | Functionalized nanoparticles and methods of use |
| CN1521231A (en) * | 2003-01-23 | 2004-08-18 | 中国人民解放军军事医学科学院微生物 | Irradiant material with surface modification and activation |
| CN102295932A (en) * | 2011-07-05 | 2011-12-28 | 武汉大学 | Method for preparing ZnSe quantum dot marked bovine serum albumin fluorescent probe |
| CN102366632A (en) * | 2011-08-22 | 2012-03-07 | 长春工业大学 | Paramagnetic metal complex functionalized fluorogold nano-cluster magnetic resonance and fluorescence imaging contrast agent |
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