CN103820106A - Green synthetic method of fluorescence copper nano-cluster, and application of fluorescence copper nano-cluster - Google Patents
Green synthetic method of fluorescence copper nano-cluster, and application of fluorescence copper nano-cluster Download PDFInfo
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- CN103820106A CN103820106A CN201410060246.8A CN201410060246A CN103820106A CN 103820106 A CN103820106 A CN 103820106A CN 201410060246 A CN201410060246 A CN 201410060246A CN 103820106 A CN103820106 A CN 103820106A
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Abstract
The invention discloses a green synthetic method of a fluorescence copper nano-cluster, and an application of the fluorescence copper nano-cluster. The synthetic method concretely comprises the following steps: incubating a certain concentration of a copper salt solution and different kinds of tumor cells, and carrying out in-situ synthesis to obtain a copper nano-cluster having an excellent fluorescence performance in order to realize the real time and high resolution fluorescence imaging of the tumor cells. The copper nano-cluster can realize the rapid real-time in-situ in-vivo tumor targeting fluorescence imaging on a tumor-hearing nude mouse model. The copper nano-cluster is prepared through direct biosynthesis of tumor cells and has a very good biocompatibility; and the method has the characteristics of environmental protection, no pollution to the environment, realization of rapid and accurate positioning and tumor targeting imaging analysis by the in-situ synthesis reaction, simplicity, easy implementation, accuracy and high efficiency.
Description
Technical field
The present invention relates to a kind of green synthesis method and application thereof of fluorescence copper nano-cluster, specifically certain density copper salt solution and different sorts tumour cell are hatched, the synthetic copper nano-cluster with excellent fluorescence property of original position, can realize real-time, the high-resolution fluorescence imaging to tumour cell thus.Meanwhile, copper nano-cluster of the present invention on model of nude mice bearing tumor, can realize fast, real-time in-situ vivo tumor target fluorescence imaging.Copper nano-cluster of the present invention is to make by the direct biosynthesizing of tumour cell, has good biocompatibility; The method environmental protection, does not pollute environment, and its in-situ synthesized reaction can be realized fast accurate location and cancer target imaging analysis, has the simple and easy to do accurately feature such as efficient.
Background technology
In life science, Biological imaging information often most important, the most directly study evidence, sometimes or even unique evidence.Be accompanied by deepening continuously of biomedical technology research, visual bio-imaging technology is being played the part of more and more important role in life science and medical field, than other bio-imaging technology, the feature such as that fluorescence imaging has is cheap, Imaging fast, sensitivity height.In addition, Imaging-PAM can be measured the tumor growth situation of various cancer models, the variation of lesion tissue in Real-Time Monitoring cancer therapy; Quantitative carries out the detection without wound ground to mouse entirety primary tumor, metastatic tumor and Spontaneous Tumor.But use at present fluorescence dye and quantum dot to have some obvious shortcomings as fluorescent probe, biological tissue as possible in lower light penetration depth destroys, and the property such as the autofluorescence of biological sample has limited its further application in bio-imaging field.
Be accompanied by Nanometer scale science and technology developed, nano material has obtained people's extensive concern for early diagnosis of cancer imaging and treatment.Nano material, because its particle is less than cancer cells, is more easily passed through barrier cell, and enhanced permeability and retention effect hyperfunction due to tumor tissues microvascular permeability and that unsound lymphatic drainage system produces, and it is preferentially assembled at tumor locus.Therefore, nano material can provide the highly sensitive of cancer patient and specificity image-forming information, can also transport the position of cancer therapy drug arrival tumour.Current, we remain limited in the understanding of the following aspects: the one, be applicable to the biomarker for imaging; The 2nd, the selection of imaging target and contrast strongthener; The 3rd, be used for the chemical process that the probe of imaging is bioid.We run into equally a lot of difficulties on developing cancer specificity imaging agents, comprising: the transportation of the probe of target tissue or tumour is not good; Bio-toxicity is large; The stability of probe is not good; The interior signal enhancing of body intensity is low etc.The present invention proposes a kind of green synthesis method and application thereof of fluorescence copper nano-cluster, specifically certain density copper salt solution and different sorts tumour cell are hatched, the synthetic copper nano-cluster with excellent fluorescence property of original position, can realize real-time, the high-resolution fluorescence imaging to tumour cell thus.
Summary of the invention
Technical scheme of the present invention provides a kind of green synthesis method and application thereof of fluorescence copper nano-cluster, specifically certain density copper salt solution and different sorts tumour cell are hatched, the synthetic copper nano-cluster with excellent fluorescence property of original position, can realize real-time, the high-resolution fluorescence imaging to tumour cell thus.Meanwhile, copper nano-cluster of the present invention on model of nude mice bearing tumor, can realize fast, real-time in-situ vivo tumor target fluorescence imaging.Copper nano-cluster of the present invention is to make by the direct biosynthesizing of tumour cell, has good biocompatibility; The method environmental protection, does not pollute environment, and its in-situ synthesized reaction can be realized fast accurate location and cancer target imaging analysis, has the simple and easy to do accurately feature such as efficient.
For the limitation of currently available technology, the invention provides a kind of green synthesis method and application thereof of fluorescence copper nano-cluster, concrete grammar is: copper salt solution and different sorts tumour cell are hatched jointly, utilize the synthetic fluorescence copper nano-cluster of specificity original position of tumour cell, realized real-time, the high-resolution fluorescence imaging to tumour cell.
Concrete measure is as follows:
First study at cell levels, its concrete steps are:
1) be that copper salt solution and the tumour cell of 0.0001mmol/L ~ 1 mmol/L hatched after 8 ~ 24 hours in cell culture incubator by concentration, obtain the synthetic copper nano-cluster of biology in situ.
2) with fluorescence spectrophotometer, confocal fluorescent microscope etc., to copper nano-cluster, the distribution situation in cell characterizes.Fluorescence distribution situation and its fluorescence intensity by fluorescence imaging are carried out qualitative or quantitative analysis to copper nano-cluster component.
When this imaging is used for vivo tumor imaging, copper salt solution is expelled to tumor tissues around or in tumor tissues, utilizes a large amount of copper nano-cluster that in tumour cell, specificity generates, use living body fluorescent imager to carry out rapid fluorescence imaging to tumor locus.Its concrete steps are:
1) build nude mice tumor model;
2) be the copper salt solution of 0.1 ~ 100 mmol/L by concentration aseptic 0.1 ~ 0.5 mL, be expelled on nude mice tumor model, on nude mice tumor model, realize real-time cancer target high resolution fluorescence imaging through 2 ~ 24 hours be incubated in; Described step 2) in injecting method be tail vein injection or local injection;
3) with living body fluorescent imager, tumor locus is carried out tumour fluorescence imaging and it carried out qualitative and and to fluorescence intensity quantitative analysis.
The present invention, compared with art methods, has the following advantages and effect:
This research adopts the method for the nanometer bio probes such as tumour growth in situ copper nano-cluster, harm and bio-toxicity that this method can effectively avoid the chemical reagent introduced in traditional nano material building-up process and nano material stablizer to cause, and can realize the analysis of vivo tumor target fluorescence imaging.
This invention is further combined with fluorescence, Raman, ultrasonic, CT and nuclear-magnetism etc., can carry out polymorphic and multi-modal synchronous imaging and accurate target location, has the simple and easy to do accurately feature such as efficient.
Accompanying drawing explanation
To describe now as preferred but nonrestrictive embodiment of the present invention, these and other features of the present invention, aspect and advantage will become apparent when read as follows detailed description with reference to accompanying drawing, wherein:
Fig. 1 is example 1 experimental group Laser Scanning Confocal Microscope result figure;
Fig. 2 is example 1 control group Laser Scanning Confocal Microscope result figure.
Embodiment
The following description is only exemplary and be not in order to limit the disclosure, application or purposes in essence.
The green synthesis method of described fluorescence copper nano-cluster and being applied as: copper salt solution and tumour cell are educated altogether, utilize a large amount of copper nano-cluster that in tumour cell, specificity generates, by fluorescence distribution situation and its fluorescence intensity of fluorescence imaging, copper nano-cluster component is carried out to qualitative or quantitative analysis.
The method of described vivo tumor imaging is: copper salt solution is expelled to around tumor tissues or in tumor tissues, utilize a large amount of copper nano-cluster that in tumour cell, specificity generates, adopt living body fluorescent imager that tumor locus is carried out tumour fluorescence imaging and it is carried out qualitative and quantitative analysis.
The present invention adopts following technical measures:
First carry out tumour cell imaging research at cell levels, its concrete steps are:
1) copper salt solution that is 0.0001mmol/L ~ 1mmol/L by concentration and tumour cell were hatched after 8 ~ 24 hours in cell culture incubator, obtained the synthetic copper nano-cluster of biology in situ.
2) with fluorescence spectrophotometer, confocal fluorescent microscope etc., to copper nano-cluster, the distribution situation in cell characterizes.Probe into the copper nano-cluster of growth in situ in tumour cell for the fluorescence imaging of the target of tumour cell with fluorescent microscope; Fluorescence distribution situation and its fluorescence intensity by fluorescence imaging are carried out qualitative or quantitative analysis to cellularstructure or chemical composition.
Described copper salt solution is the one in title complex or the salt of the copper such as cupric nitrite, copper sulfate, cupric chloride, cupric phosphate, neutralized verdigris.
Described tumour cell is the tumor cell lines such as liver cancer, lung cancer, cervical cancer, leukemia, osteosarcoma.
The method of tumour cell imaging confirm copper salt solution can with the synthetic copper nano-cluster of tumour cell biology in situ, on this basis, copper salt solution is injected to vivo tumor cell tissue around or tumor tissues is realized living body fluorescent imaging.
Study at living animal model level, its concrete steps are:
1) build nude mice tumor model;
2) be the copper salt solution of 0.1 ~ 100 mmol/L by concentration aseptic 0.1 ~ 0.5 mL, be expelled on nude mice tumor model, on nude mice tumor model, realize quick in situ, cancer target high resolution fluorescence imaging in real time through 2 ~ 24 hours be incubated in;
3) utilize process 2) in the copper nano-cluster that generates, use Imaging-PAM to carry out fluorescence imaging to tumor locus, thereby distribution and the quantity of the biochemical composition to tumor locus are carried out qualitative and quantitative analysis according to glimmering light intensity and distribution situation, to realize early diagnosis and the tumor therapeutic procedure timing monitoring of tumour.Described step 2) in injecting method be tail vein injection or local injection.
example 1based on method and the application thereof of the synthetic copper nano-cluster of biology in situ in tumour cell
1. take HepG2 cell as research object, by the aseptic copper salt solution for 0.0001mmol/L ~ 1 mmol/L with hatch altogether 8 ~ 24 hours (37 ° of C, 5 % CO in the HepG2 of logarithmic phase cell
2, RH 95%), can obtain the synthetic copper nano-cluster of biology in situ in cell paste.
2. with confocal fluorescent microscope etc., copper compound nano-cluster is carried out to qualitative and quantitative analysis;
Select liver cancer cell (HepG2) as research object, experimental group by the liver cancer cell in logarithmic phase (HepG2) according to 1.6 × 10
5the density of individual cells/well is inoculated in 6 orifice plates, cultivate add after 24 h sterilizing and dilute with the DMEM substratum of fresh sterile contain the copper salt solution that concentration is 0.0001mmol/L ~ 1 mmol/L.
Control group by the liver cancer cell in substratum (HepG2) according to 1.6 × 10
5the density of individual cells/well is inoculated in 6 orifice plates, cultivates 24 h.Every hole that incubation time stops backward experimental group and control group adds phosphate buffer solution (PBS, pH=7.2) to rinse 2-3 time.Be placed under laser focusing fluorescent microscope, adopting wavelength is that 488 nm blue lights excite the green fluorescence image that can collect cell, can observe clearly this copper nano-cluster by fluorescence layer scanning technology mainly concentrates on cytolemma and tenuigenin, as shown in Figure 1 and Figure 2, it should be noted that, although picture background color is darker, examine electronic pictures still distinguishable go out, this does not affect of the present invention fully open.
example 2based on the cancer target imaging of live body focus growth in situ copper nano-cluster
First near the tumour of implanting Hepatic neoplasm model nude mice, the aseptic concentration of local subcutaneous injection 0.1 ~ 0.5 mL is the copper salt solution of 0.1 ~ 100 mmol/L, after injection, choose respectively 2 hours, the different time points such as 4 hours, 8 hours and 24 hours hatches, then this experiment nude mice is carried out to gas anesthesia with 5% isoflurane, then be placed on small animal living body imager service platform, select the blue-light excited image that can collect tumor region.
The above is only the preferred embodiment of the present invention; be noted 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 (6)
1. the synthetic application method of the green of a fluorescence copper nano-cluster, it is characterized in that, the method is hatched copper salt solution and different sorts tumour cell, utilize the synthetic copper nano-cluster of specificity original position of tumour cell, can realize thus real-time, the high-resolution fluorescence imaging to tumour cell, its concrete steps are:
1) be that copper salt solution and the tumour cell of 0.0001mmol/L ~ 1 mmol/L hatched after 8 ~ 24 hours in cell culture incubator by concentration, obtain the synthetic copper nano-cluster of biology in situ;
2) with fluorescence spectrophotometer and confocal fluorescent microscope, to copper nano-cluster, the distribution situation in cell characterizes, and by fluorescence distribution situation and its fluorescence intensity of fluorescence imaging, copper nano-cluster component is carried out to qualitative or quantitative analysis.
2. the synthetic application method of the green of fluorescence copper nano-cluster as claimed in claim 1, wherein said copper salt solution is the one in title complex or the salt of the copper such as cupric nitrite, copper sulfate, cupric chloride, cupric phosphate, neutralized verdigris.
3. the synthetic application method of the green of fluorescence copper nano-cluster as claimed in claim 1, is characterized in that: described tumour cell is the tumor cell line being selected among liver cancer, lung cancer, cervical cancer, leukemia, osteosarcoma.
4. the synthetic application method of the green of fluorescence copper nano-cluster as claimed in claim 1, it is characterized in that, when the method is used for vivo tumor imaging, copper salt solution is expelled to around tumor tissues or in tumor tissues, utilize the specificity reaction in－situ of tumor tissues to generate fluorescence copper nano-cluster, use living body fluorescent imager to carry out fluorescence imaging to tumor locus, its concrete steps are:
1) build nude mice tumor model;
2) be the copper salt solution of 0.1 ~ 100 mmol/L by concentration aseptic 0.1 ~ 0.5 mL, be expelled on nude mice tumor model, on nude mice tumor model, realize quick in situ, cancer target high resolution fluorescence imaging in real time through 4 ~ 24 hours be incubated in;
3) with living body fluorescent imager, tumor locus is carried out tumor imaging and it is carried out qualitative and quantitative analysis.
5. the synthetic application method of the green of fluorescence copper nano-cluster as claimed in claim 4, is characterized in that described step 2) in injecting method be tail vein injection or local injection.
6. the synthetic application method of the green of the fluorescence copper nano-cluster as described in claim 1-5 any one, is characterized in that, described fluorescence copper nano-cluster can be realized fast on model of nude mice bearing tumor, real-time in-situ vivo tumor target fluorescence imaging.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104972135A (en) * | 2015-05-28 | 2015-10-14 | 天津师范大学 | Synthesis method for near-infrared fluorescent probe copper nano-cluster and application of synthesis method |
| CN105486670A (en) * | 2016-01-18 | 2016-04-13 | 大连理工大学 | Detection method for divalent Co ions |
| CN107338042A (en) * | 2017-07-17 | 2017-11-10 | 吉林大学 | Full color emission copper cluster assembly fluorescent material, preparation method and its application in white light LED part of solvent regulation and control |
| CN108526485A (en) * | 2018-04-27 | 2018-09-14 | 广州大学 | A kind of preparation method and applications of the metallic copper nanocluster of no surface ligand cladding |
| CN109128217A (en) * | 2018-11-09 | 2019-01-04 | 云南大学 | A kind of preparation method of one-step synthesis method green fluorescence copper nanocluster |
| CN114280024A (en) * | 2021-12-25 | 2022-04-05 | 福州大学 | Specific nitrite fluorescence detection method based on copper nanocluster and oxidation reaction |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102703060A (en) * | 2012-06-08 | 2012-10-03 | 中国药科大学 | Targeted tracing noble metal fluorescence probe and anti-tumor prodrug |
| CN103212056A (en) * | 2013-05-07 | 2013-07-24 | 东南大学 | Preparation based on gold, silver, gold-silver mixture and glutathione/chitosan, and application of preparation |
| CN103330948A (en) * | 2013-07-05 | 2013-10-02 | 东南大学 | Tumor targeted living body rapid fluorescence imaging method of rare earth metal nanocluster |
-
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102703060A (en) * | 2012-06-08 | 2012-10-03 | 中国药科大学 | Targeted tracing noble metal fluorescence probe and anti-tumor prodrug |
| CN103212056A (en) * | 2013-05-07 | 2013-07-24 | 东南大学 | Preparation based on gold, silver, gold-silver mixture and glutathione/chitosan, and application of preparation |
| CN103330948A (en) * | 2013-07-05 | 2013-10-02 | 东南大学 | Tumor targeted living body rapid fluorescence imaging method of rare earth metal nanocluster |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104972135A (en) * | 2015-05-28 | 2015-10-14 | 天津师范大学 | Synthesis method for near-infrared fluorescent probe copper nano-cluster and application of synthesis method |
| CN104972135B (en) * | 2015-05-28 | 2017-03-01 | 天津师范大学 | The synthetic method of near infrared fluorescent probe copper nano-cluster and its application |
| CN105486670A (en) * | 2016-01-18 | 2016-04-13 | 大连理工大学 | Detection method for divalent Co ions |
| CN105486670B (en) * | 2016-01-18 | 2018-07-13 | 大连理工大学 | A kind of detection method of divalent cobalt ion |
| CN107338042A (en) * | 2017-07-17 | 2017-11-10 | 吉林大学 | Full color emission copper cluster assembly fluorescent material, preparation method and its application in white light LED part of solvent regulation and control |
| CN108526485A (en) * | 2018-04-27 | 2018-09-14 | 广州大学 | A kind of preparation method and applications of the metallic copper nanocluster of no surface ligand cladding |
| CN108526485B (en) * | 2018-04-27 | 2021-06-08 | 广州大学 | Preparation method and application of metal copper nanocluster without surface ligand coating |
| CN109128217A (en) * | 2018-11-09 | 2019-01-04 | 云南大学 | A kind of preparation method of one-step synthesis method green fluorescence copper nanocluster |
| CN114280024A (en) * | 2021-12-25 | 2022-04-05 | 福州大学 | Specific nitrite fluorescence detection method based on copper nanocluster and oxidation reaction |
| CN114280024B (en) * | 2021-12-25 | 2023-10-20 | 福州大学 | Specific nitrite fluorescent detection method based on copper nanoclusters and oxidation reaction |
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Application publication date: 20140528 |