CN103965905A - Synthetic method for folic acid-modified water-soluble rare-earth up-conversion luminescence nanocrystalline - Google Patents
Synthetic method for folic acid-modified water-soluble rare-earth up-conversion luminescence nanocrystalline Download PDFInfo
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Abstract
The invention relates to a synthetic method for a folic acid-modified water-soluble rare-earth up-conversion luminescence nanocrystalline. The synthetic method comprises the following steps: firstly, activating folic acid; then, adding the activated folic acid to a cysteine-modified rare-earth up-conversion luminescence nanocrystalline aqueous dispersion, and conducing a reaction for a certain time under the conditions of normal pressure, normal temperature and light shielding to form a folic acid-modified nanocrystalline; finally, conducting separation and purification so as to obtain the folic acid-modified water-soluble rare-earth up-conversion luminescence nanocrystalline. The folic acid-modified water-soluble fluorescent nanocrystalline material has the advantages of uniform size, stable structure, higher up-conversion fluorescence intensity, high biocompatibility and the like. The synthetic method can be applied to surface modification of nanocrystalline doped with different rare earth ions. The product can be applied to the up-conversion fluorescence imaging of hela cells, with obvious effect and good targeting. Therefore, the material has a potential biological application value in the targeting imaging of cells.
Description
Technical field
The present invention relates to the synthetic method of the water-solubility rare-earth up-conversion luminescence nanometer crystal of a kind of synthetic method of water-solubility rare-earth up-conversion luminescence nanometer crystal, particularly a kind of modified with folic acid.
Background technology
Optical imagery with its Non-Invasive, in real time, resolving power is high etc., and advantage is widely used in tumor research field, can carry out early diagnosis to tumour, reflection tumour anatomical structure and metabolism situation.Fluorescence imaging is one of focus of current optical molecular imaging area research, and it can the biological esoteric physiological process of direct-detection and pathological state.Rear-earth-doped up-conversion luminescence nano material is emerging in the last few years a kind of fluorescent material, it is utilize near infrared light to excite and send visible ray or near infrared light, this characteristic has overcome the deficiency of the poor and poor signal to noise of conventional fluorescent material light penetrativity, can be advantageously applied to Biological Detection, and rear-earth-doped up-conversion nano material synthesis technique is simple, and pattern size is controlled.
Targeted drug is the high-tech medicine producing along with the development of contemporary molecular biology, cytobiology, the current state-of-the-art medicine that is used for the treatment of cancer, it by occurring with cancer, the growth that is used for stoping cancer cells of the necessary specific molecular target spot of tumor growth.And targeted drug is developed for oncogene, the characteristic site that it is determined by the distinctive gene of tumour cell on can tumor cell, by with it in conjunction with (or similarly other mechanism), in blocking-up tumour cell, control the signal transduction pathway of Growth of Cells, propagation, thereby killing off tumor cells, stop its propagation.Due to like this, targeted drug is not only effective, and side effect is more much smaller than conventional embolic chemotherapy.
Summary of the invention
The object of the invention is to the synthetic method of the water-solubility rare-earth up-conversion luminescence nanometer crystal that proposes a kind of modified with folic acid, prepared luminescence nanocrystalline has good targeted imaging effect to HeLa cell.
For achieving the above object, technical scheme provided by the present invention is:
A synthetic method for the water-solubility rare-earth up-conversion luminescence nanometer crystal of modified with folic acid, is characterized in that the concrete steps of the method are:
A. by folic acid, 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and N-succinic acid, S-ethanoyl sulfydryl glycol ester (NHS) is dissolved in deionized water by the mass ratio of 2:1.5 ~ 2.5:0.5 ~ 1, and the concentration of controlling folic acid is 0.3 ~ 0.6 mg/mL; After mixing, under lucifuge condition, react 2 ~ 5 hours, make the first dispersion liquid;
B. the rare earth up-conversion luminescence nanometer crystal of halfcystine modification is added in deionized water to ultrasonic dispersion, the second dispersion liquid that formation concentration is 2.5～6mg/mL;
C. first, second dispersion liquid is mixed by the volume ratio of 2.5 ~ 3.5:1, react 12 hours under lucifuge condition, centrifugation, makes the water-solubility rare-earth up-conversion luminescence nanometer crystal of modified with folic acid.
The inventive method is a kind of synthetic method of water-solubility rare-earth up-conversion luminescence nanometer crystal of modified with folic acid specifically.First, folic acid is activated; Then joined in the rare earth upconversion nano crystal aqueous dispersions of halfcystine modification, and reacted the regular hour under normal temperature and pressure lucifuge condition, formed the nanocrystalline of modified with folic acid; Finally, be isolated purifying, obtain the water-solubility rare-earth up-conversion luminescence nanometer crystal of modified with folic acid.It is simple that the inventive method has technique, easy to operate, the advantage of easy control of structure.Not only size homogeneous, the Stability Analysis of Structures of nano-luminescent material of modifying, and there is the advantages such as up-conversion fluorescence is strong, good biocompatibility, and can be used for the surface modification of different rare earth ions Doped nanocrystal, product can be applied to the target up-conversion fluorescence imaging of HeLa cell.
Brief description of the drawings
Fig. 1 is the TEM photo of the embodiment of the present invention 1 gained sample;
Fig. 2 is the fluorescence spectrum figures of the embodiment of the present invention 3 gained samples under 980 nm LASER Light Source excite;
Fig. 3 is the cell imaging picture of application examples gained sample of the present invention.
Embodiment
Below in conjunction with drawings and Examples, the present invention is described in detail:
embodiment 1:
Preparation method's reference [1] of water-solubility rare-earth up-conversion luminescence nanometer crystal.
The present embodiment provides a kind of synthetic method of water-solubility rare-earth up-conversion luminescence nanometer crystal of modified with folic acid, it is characterized in that the method comprises following processing step:
A. by folic acid activation, form the first dispersion liquid;
By folic acid, 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and N-succinic acid, S-ethanoyl sulfydryl glycol ester (NHS) is dissolved in deionized water by the mass ratio of 2:1.5 ~ 2.5:0.5 ~ 1, and the concentration of controlling folic acid is 0.3 ~ 0.6 mg/mL; After mixing, under lucifuge condition, react 2 ~ 5 hours, make the first dispersion liquid;
B. by the rare earth up-conversion luminescence nanometer crystal NaYF of halfcystine modification
4: Yb, Er-Cys adds in deionized water, ultrasonic dispersion, the second dispersion liquid that formation concentration is 2.5～6mg/mL;
C. first, second dispersion liquid is mixed by the volume ratio of 2.5 ~ 3.5:1, under lucifuge condition, react 12 hours, generate the nanometer product of modified with folic acid;
D. by the centrifugation of nanometer product, make the water-solubility rare-earth up-conversion luminescence nanometer crystal NaYF of the modified with folic acid to HeLa cell target
4: Yb, Er-FA;
embodiment 2:
The present embodiment provides a kind of synthetic method of water-solubility rare-earth up-conversion luminescence nanometer crystal of modified with folic acid, and it comprises the following steps:
A. by folic acid activation, form the first dispersion liquid;
A. by folic acid activation, form the first dispersion liquid;
By folic acid, 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and N-succinic acid, S-ethanoyl sulfydryl glycol ester (NHS) is dissolved in deionized water by the mass ratio of 2:1.5 ~ 2.5:0.5 ~ 1, and the concentration of controlling folic acid is 0.3 ~ 0.6 mg/mL; After mixing, under lucifuge condition, react 2 ~ 5 hours, make the first dispersion liquid;
B. by the rare earth up-conversion luminescence nanometer crystal NaYF of halfcystine modification
4: Yb, Tm-Cys adds in deionized water, ultrasonic dispersion, the second dispersion liquid that formation concentration is 2.5～6mg/mL;
C. first, second dispersion liquid is mixed by the volume ratio of 2.5 ~ 3.5:1, under lucifuge condition, react 12 hours, generate the nanometer product of modified with folic acid;
D. by the centrifugation of nanometer product, make the water-solubility rare-earth up-conversion luminescence nanometer crystal NaYF of the modified with folic acid to HeLa cell target
4: Yb, Tm-FA;
embodiment 3:
The present embodiment provides a kind of synthetic method of water-solubility rare-earth up-conversion luminescence nanometer crystal of modified with folic acid, and it comprises the following steps:
A. by folic acid activation, form the first dispersion liquid;
By folic acid, 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and N-succinic acid, S-ethanoyl sulfydryl glycol ester (NHS) is dissolved in deionized water by the mass ratio of 2:1.5 ~ 2.5:0.5 ~ 1, and the concentration of controlling folic acid is 0.3 ~ 0.6 mg/mL; After mixing, under lucifuge condition, react 2 ~ 5 hours, make the first dispersion liquid;
B. by the rare earth up-conversion luminescence nanometer crystal NaYF of halfcystine modification
4: Yb, Er, Tm-Cys adds in deionized water, ultrasonic dispersion, the second dispersion liquid that formation concentration is 2.5～6mg/mL;
C. first, second dispersion liquid is mixed by the volume ratio of 2.5 ~ 3.5:1, under lucifuge condition, react 12 hours, generate the nanometer product of modified with folic acid;
D. by the centrifugation of nanometer product, make the water-solubility rare-earth up-conversion luminescence nanometer crystal NaYF of the modified with folic acid to HeLa cell target
4: Yb, Er, Tm-FA;
application example:
The present embodiment provides the water-solubility rare-earth up-conversion luminescence nanometer crystal of modified with folic acid to be applied to HeLa cell imaging experiment, and it comprises the following steps:
A. for cell imaging;
A1. prepare the water-solubility rare-earth up-conversion luminescence nanometer crystal of 10 mg modified with folic acid, 5 mL physiological saline, cell culture fluid, grows and has 1 of the slide glass of 1 of the slide glass of HeLa cell, MCF-7 cell;
A2. the water-solubility rare-earth up-conversion luminescence nanometer crystal of the modified with folic acid of preparation, physiological saline and cell culture fluid water are mixed, within ultrasonic 10 minutes, disperse, form the 3rd dispersion liquid;
A3. get the 3rd dispersion liquid 1mL and add in the culture dish that slide glass is housed, cultivate 3 hours with being placed in constant temperature and humidity machine case, under laser confocal microscope, observe.
Can find out by Fig. 1 to Fig. 3,
Fig. 1 is the water-solubility rare-earth up-conversion luminescence nanometer crystal NaYF of the embodiment of the present invention 1 gained sample modified with folic acid
4: Yb, the TEM photo of Er-FA, as can be seen from the figure nano particle diameter is evenly distributed, with raw material NaYF
4: Yb, Er-Cys compares size and does not change, and nanoparticle median size after modification is about 30 nm, because small size nanoparticle is more easily by cell endocytic, this circulates significant for it as bioprobe in vivo.
Fig. 2 is the water-solubility rare-earth up-conversion luminescence nanometer crystal NaYF of the embodiment of the present invention 3 gained sample modified with folic acid
4: Yb, Er, the fluorescence spectrum figure of Tm-FA under 980 nm LASER Light Source excite, is positioned at 521 nm as can be observed from Figure, 542 nm, the emission peak at 654 nm places, corresponds respectively to Er
3+'s
2h
11/2→
4i
15/2,
4s
3/2→
4i
15/2,
4f
9/2→
4i
15/2the transition of energy level; And 475 nm, the emission peak at 802 nm places, corresponds respectively to Tm
3+'s
1g
4→
3h
6,
3h
4→
3h
6the transition of energy level, shows that the up-conversion nano material after modified with folic acid still keeps good fluorescence property.
Fig. 3 is the water-solubility rare-earth up-conversion luminescence nanometer crystal NaYF of application example gained modified with folic acid of the present invention
4: Yb, Er, the cell imaging photo of Tm-FA, wherein a is Hela cell imaging photo, and b is MCF-7 cell photo, contrasts known by a, b picture, the water-solubility rare-earth up-conversion luminescence nanometer crystal of modified with folic acid is better to the Targeting Effect of Hela cell, and fluorescence intensity is stronger, signal to noise ratio is large, illustrates that this nano material is the excellent nano material to the imaging of Hela cell-targeting.
According to the synthetic method of the water-solubility rare-earth up-conversion luminescence nanometer crystal of modified with folic acid provided by the present invention, the product obtaining is size homogeneous, Stability Analysis of Structures not only, and there is the advantages such as up-conversion fluorescence is stronger, particularly product can be applicable to the target up-conversion fluorescence imaging of HeLa cell, and successful.
The inventive method has simple, the easy to operate advantage of technique, specifically a kind of synthetic and application of water-solubility rare-earth up-conversion luminescence nanometer crystal of modified with folic acid.First, folic acid is activated; Then joined in the rare earth upconversion nano crystal aqueous dispersions of halfcystine modification, and reacted the regular hour under normal temperature and pressure lucifuge condition, formed the nanocrystalline of modified with folic acid; Finally, be isolated purifying, obtain the rare earth up-conversion luminescence nanometer crystal of modified with folic acid.Not only size homogeneous, Stability Analysis of Structures of gained nano-luminescent material of the present invention, and there is the advantages such as up-conversion fluorescence is strong, good biocompatibility, and the nanocrystalline folic acid surface modification that can be used for different rare earth ions doping, product can be applied to the target up-conversion fluorescence imaging of HeLa cell.
reference:
35 (2014) 387-392。
Claims (1)
1. a synthetic method for the water-solubility rare-earth up-conversion luminescence nanometer crystal of modified with folic acid, is characterized in that the concrete steps of the method are:
A. by folic acid, 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and N-succinic acid, S-ethanoyl sulfydryl glycol ester (NHS) is dissolved in deionized water by the mass ratio of 2:1.5 ~ 2.5:0.5 ~ 1, and the concentration of controlling folic acid is 0.3 ~ 0.6 mg/mL; After mixing, under lucifuge condition, react 2 ~ 5 hours, make the first dispersion liquid;
B. the rare earth up-conversion luminescence nanometer crystal of halfcystine modification is added in deionized water to ultrasonic dispersion, the second dispersion liquid that formation concentration is 2.5～6mg/mL;
C. first, second dispersion liquid is mixed by the volume ratio of 2.5 ~ 3.5:1, react 12 hours under lucifuge condition, centrifugation, makes the water-solubility rare-earth up-conversion luminescence nanometer crystal of modified with folic acid.
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| CN104804739A (en) * | 2015-03-27 | 2015-07-29 | 上海大学 | Preparation method of POSS (polyhedral oligomeric silsesquioxane) modified up-conversion nanocrystalline |
| CN111995759A (en) * | 2020-02-15 | 2020-11-27 | 江西师范大学 | Rare earth-folic acid coordination polymer nano particle and preparation method thereof |
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Application publication date: 20140806 |