CN111518558B - C3N4Nanosphere loaded all-inorganic perovskite CsPbBr3Preparation method of (1) and electrochemiluminescence cell sensing thereof - Google Patents
C3N4Nanosphere loaded all-inorganic perovskite CsPbBr3Preparation method of (1) and electrochemiluminescence cell sensing thereof Download PDFInfo
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Abstract
The invention belongs to the field of biological analysis and electrochemiluminescence, and particularly relates to C3N4Nanosphere loaded all-inorganic perovskite CsPbBr3The preparation method and the electrochemiluminescence cell sensing thereof, and the C is prepared by an in-situ growth method3N4Nanosphere loaded CsPbBr3The composite material is used as an electrode light-emitting substrate; then designing a probe which comprises a target recognition part and a multifunctional DNA part, wherein the multifunctional DNA part also comprises an enzyme cutting site, a hybridization recognition fragment and a hybridization chain reaction initiation fragment; the probe firstly targets a cell surface receptor specifically, then a multifunctional DNA part of the probe is cut by using a restriction endonuclease Dra I, and a hybridization chain reaction is initiated in a solution to generate a large number of long DNA chains for embedding rhodamine 6G; the product is then captured to the electrode surface by hybridization; due to CsPbBr in the composite material3Can generate efficient resonance energy transfer with rhodamine 6G to cause the reduction of electrochemiluminescence signals of the rhodamine, thereby realizing the analysis of the cell number.
Description
Technical Field
The invention belongs to the field of biological analysis and electrochemiluminescence, and particularly relates to C3N4Nanosphere loaded all-inorganic perovskite CsPbBr3The preparation method and the electrochemiluminescence cell sensing thereof.
Background
The research on semiconductor nano materials is one of the hot spots of current material science research, and the unique optical and electrical properties of the semiconductor nano materials enable the semiconductor nano materials to be widely applied in various fields. Further improving the performance of the nano material and developing the unique photoelectric property of the novel nano material, so that the novel nano material can be better applied to practical problems, and is always one of the concerns of researchers. In recent years, lead perovskite halide has been widely used as a brand new semiconductor material in the fields of solar cells, light emitting diodes, lasers, photodetectors, capacitors, photocatalysis, and the like (y.cao, z.zhang, l.li, j.r.zhang, j.j.zhu, anal.chem.2019,91,8607). They have excellent spectral and optoelectronic properties, such as high photoluminescence quantum yield, narrow emission bandwidth, tunable band gap, strong optical absorption capability, low exciton binding energy, high charge carrier mobility and long carrier lifetime (k.lin, j.xing, l.n.quan, f.p.g.de Arquer, x.gong, j.lu, l.xie, w.zhao, d.zhang, c.yan, w.li, x.liu, y.lu, j.kiran, e.h.sargent, q.xiong, z.wei, Nature 2018,562,245), among which all-inorganic perovskites (CsPbX, x.x, j.k.r.j.k.r.3X ═ Cl, Br, I) exhibit inter alia surprising properties, including strong and narrow fluorescence emission, excellent thermal stability, and low humidity sensitivity. These desirable properties suggest that all-inorganic perovskite materials can be extended to many other optically and electrically related fields.
Electrochemiluminescence describes the phenomenon of photoemission of excited species triggered by the modulated potential at the electrode surface during energy relaxation, which is a combination of electricity and optics. Theoretically, all-inorganic perovskite materials appear to be tailored electrochemiluminescent emitters due to their optoelectronic properties. Then until recently its electrochemiluminescence phenomenon was reported, CsPbBr3Nanocrystals can obtain ECL signals through self-annihilation or co-reaction mechanisms(Y.Huang, M.Fang, G.Zou, B.Zhang, H.Wang, Nanoscale 2016,8, 18734; J.Xue, Z.Zhang, F.ZHEN, Q.Xu, J.Xu, G.Zou, L.Li, J.J.Zhu, anal.chem.2017,89,8212.). Unfortunately, these electrochemiluminescence processes can only be achieved in organic systems due to their inherent vulnerability to water. Therefore, the water phase property of deep-level all-inorganic perovskite nano-materials has not been paid attention, and the electrochemical redox property, the electrochemiluminescence mechanism and the surface state of nano-crystals in aqueous solution are still to be further researched. The electrochemiluminescence properties are poor due to its intrinsic affinity to moisture, and are easily destroyed, hindering their potential applications under normal conditions (w.ke, m.g. kantzidis, nat. commun.2019,10,965; a. louudice, s.saris, e.oveisi, d.t.l.alexander, r.buonsani, angelw.chem.int.ed.2017, 56, 10696.). Unfortunately, reliable electrochemiluminescence applications of all-inorganic perovskite nanomaterials have not been achieved to date.
Disclosure of Invention
The invention solves the technical problems in the prior art and provides a C3N4Nanosphere loaded all-inorganic perovskite CsPbBr3The method for synthesizing the composite material, and an analysis method for detecting the number of cells based on electrochemiluminescence of the composite material.
In order to solve the problems, the technical scheme of the invention is as follows:
c3N4Nanosphere loaded all-inorganic perovskite CsPbBr3The method for synthesizing the composite material comprises the following steps: the method comprises the following steps:
(1) 0.2035g of Cs are taken2CO3Mixing 5-10 mL of octadecene and 0.5-0.625 mL of oleic acid, and then drying in vacuum; then heating the mixture in an inert gas atmosphere until the solid is completely dissolved to form a precursor solution;
(2) 0.138g of PbBr was taken21-5 mg of hollow mesoporous C3N4Mixing the nanospheres with 5-10 mL of octadecene, and then drying in vacuum; adding oleylamine and oleic acid, and heating to 120-150 ℃ in an inert gas atmosphere to form a high-temperature reaction solution;
(3) adding 1.0mL of the precursor solution prepared in the step (1) into the high-temperature reaction solution in the step (2), wherein the volume ratio of the precursor solution to the high-temperature reaction solution is as follows: 1: 5-10; and after reacting for 3-5 s, placing the mixture in an ice water bath for cooling to terminate the reaction, and obtaining a crude product.
Preferably, the crude product obtained in step (3) is worked up by centrifuging, collecting the centrifuged solid, washing, dispersing in toluene and storing for later use. The centrifugation conditions are preferably 8000rpm for 10 min.
Preferably, the step (1) is heated to 150 ℃ in an inert gas atmosphere until the solid is completely dissolved to form the precursor solution.
Preferably, the vacuum drying conditions of the step (1) and the step (2) are as follows: placing in a vacuum drying oven at 100 deg.C for 1 h.
Preferably, the inert gas atmosphere of the step (1) and the step (2) is N2And (4) atmosphere.
Preferably, in the step (2), the oleylamine and the oleic acid are dried in a vacuum oven at 100 ℃ for 1 hour in advance.
Preferably, in the step (2), the temperature is increased to 120 ℃ in an inert gas atmosphere and is kept for 40min, and then the heating is continued to 150 ℃ to form a high-temperature reaction solution.
Based on C3N4Nanosphere loaded all-inorganic perovskite CsPbBr3The analysis method for detecting the cell number by electrochemiluminescence of the composite material comprises the following steps:
a) the preparation process of the electrode comprises the following steps: with said C3N4Nanosphere loaded all-inorganic perovskite CsPbBr3The glassy carbon electrode is sequentially modified by capturing DNA;
b) the preparation process of the probe comprises the following steps: equimolar mixing of the connecting DNA and the functional DNA, and placing the mixture in a constant-temperature oscillator at 37 ℃ for reaction; then adding a target recognition molecule, 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide and N-hydroxysuccinimide to react in a constant temperature oscillator at 37 ℃; preparing a probe by centrifugal purification;
c) evaluation process of cell surface receptors: incubating the probe prepared in the step b) and the cells in an incubator at 37 ℃, centrifuging, collecting precipitates, washing, and dispersing in a phosphate buffer solution with the pH value of 7.4; then adding restriction enzyme DraI to specifically cut DNA probes on the cell surface in a thermostat (37 ℃) for 40min, centrifuging and collecting supernate, respectively adding hairpin DNA1 and hairpin DNA2, and placing the supernate in a thermostat-controlled oscillator at 37 ℃ for carrying out hybridization chain reaction; finally adding rhodamine 6G, and reacting at room temperature;
d) electrochemiluminescence detection process: dropwise adding the reaction product formed in the step c) to the surface of the electrode prepared in the step a), reacting at room temperature, and measuring an electrochemiluminescence signal; with constant C of the cathode3N4Electrochemiluminescence signal of spheres as internal standard, anode-changed CsPbBr3The electrochemiluminescence signal of (a) as an indicator signal, an analytical strategy for internal reference is achieved by the ratio of the two signals, which ratio exhibits a good linear relationship with the cell concentration.
The targeting molecule in step b) specifically targets the cell surface receptor of step c).
Preferably, in step b), the functional DNA portion comprises an enzyme cleavage site fragment, a hybridization recognition fragment, and a priming fragment for hybridization chain reaction.
Preferably, the electrochemiluminescence measurement of step d) is performed in a three-electrode system in a phosphate buffer solution containing 1mM ascorbic acid at pH 7.4, including the prepared electrode as a working electrode, a platinum wire as a counter electrode, and a saturated calomel electrode as a reference electrode.
Preferably, in step d), the electrochemiluminescence signals are all measured by a photomultiplier under the same high pressure condition.
Compared with the prior art, the invention has the advantages that,
the invention successfully synthesizes C with excellent stability and high electrochemiluminescence efficiency3N4Nanosphere loaded all-inorganic perovskite CsPbBr3The composite material and the application of electrochemiluminescence biological analysis to the composite material realize the evaluation of cell surface receptors.
Has the following advantages:
(1) by mixing CsPbBr3Encapsulation of nanocrystals into C3N4Inside the nanosphere, not only CsPbBr is improved3The material is unstable, the electrochemiluminescence performance is obviously improved, and CsPbBr is realized for the first time3The application of the nano crystal in electrochemiluminescence biological analysis;
(2) the dual signal amplification strategy improves the sensitivity of cell surface receptor assessment: one is signal amplification of the probe itself, since a single targeting molecule can assemble multiple functional DNAs; secondly, signal amplification based on chain hybridization reaction;
(3)CsPbBr3the electrochemiluminescence of the nano crystal has the characteristic of narrow half-peak width, and is superior to all reported electrochemiluminescence emitters at present; therefore, an electrochemiluminescence resonance energy transfer system constructed by utilizing the property shows extremely high transfer efficiency, so that the sensitivity of an analysis process is further increased;
(4) with C3N4Electrochemiluminescence signal of nanospheres as internal standard, CsPbBr3The electrochemiluminescence signal of the nano crystal is used as an indicating signal, and the accuracy of the detection process is improved by an internal reference analysis method.
Drawings
FIG. 1 shows a graph of C prepared according to the present invention3N4Nanospheres (A) and C3N4Nanosphere loaded all-inorganic perovskite CsPbBr3Transmission electron microscopy characterization of the composite material (B);
FIG. 2 is a schematic diagram of the present invention relating to the evaluation process of MCF-7 cell surface CD44 receptor;
FIG. 3 is a graph showing the actual electrochemiluminescence detection curves (A) of MCF-7 cells overexpressing CD44 analyzed by the present invention, wherein a-h represent MCF-7 cell concentrations of 3.2X 10, respectively2,103,3.2×103,104,3.2×104,105,3.2×105,106cells/mL; linear plot of MCF-7 cell concentration versus electrochemiluminescence intensity for CD44 over-expressed (B).
Detailed Description
Example 1:
the invention provides a method based on C3N4Nanosphere loaded all-inorganic perovskite CsPbBr3The bioanalytical method of detecting the concentration of MCF-7 cells overexpressing CD 44.
C3N4Nanosphere loaded all-inorganic perovskite CsPbBr3The method for synthesizing the composite material comprises the following steps: 0.2035g of Cs are taken2CO310mL of octadecene and 0.625mL of oleic acid are respectively added into a three-neck flask and placed in a vacuum drying oven at 100 ℃ for 1 h; then in N2The atmosphere was heated to 150 ℃ until the solid was completely dissolved to form a precursor solution. 0.138g of PbBr was taken21mg of hollow mesoporous C3N4Respectively adding the nanospheres (figure 1A) and 10mL of octadecene into a three-neck flask, and placing the three-neck flask in a vacuum drying oven at 100 ℃ for 1 h; followed by 1.0mL of oleylamine and 1.0mL of oleic acid, previously dried at 100 ℃ under vacuum, and then added under N2Heating to 120 deg.C in atmosphere, maintaining for 40min, and further heating to 150 deg.C to form high temperature reaction solution. And (3) quickly injecting 1.0mL of precursor solution into the high-temperature reaction solution, reacting for 5s, and then placing the solution in an ice water bath to cool and terminate the reaction. The reaction crude product was centrifuged at 8000rpm for 10min, the centrifuged solid was collected and washed with ethyl acetate and toluene once, and the finally collected solid was redispersed in toluene for storage and its characterization by transmission electron microscopy is shown in FIG. 1.
Detection of the concentration of MCF-7 cells overexpressing CD44 (see FIG. 2): mixing 10. mu.L of the above C3N4Nanosphere loaded all-inorganic perovskite CsPbBr3The composite material is dripped on the surface of a glassy carbon electrode; after natural drying, 10 mu L of capture DNA is dripped; after 1h, dropwise adding 10 mu L of mercaptohexanol to seal the residual active sites on the surface of the material; finally, the mixture is washed clean by distilled water and is washed in N2Drying in the atmosphere. Equimolar mixing connecting DNA with amino at the 5' end and functional DNA, and placing the mixture in a constant-temperature oscillator at 37 ℃ for reaction for 4 hours; then adding hyaluronic acid, 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide (EDC) and N-hydroxysuccinimide (NHS) to react for 5 hours in a constant temperature shaking instrument at 37 ℃; finally, the probe is purified by centrifuging for many times by using an ultrafiltration tube with 3KD. Incubating MCF-7 cells with 0.5 μ M probe for 1h at 37 ℃, centrifuging at 1000rpm for 5min, and washing the collected centrifugation pellet with phosphate buffer (pH 7.4); after three repetitions, the final centrifugation pellet was dispersed in phosphate buffer (pH 7.4); then adding restriction enzyme DraI to specifically cut DNA probes on the cell surface in a thermostat (37 ℃) for 40min, centrifuging and collecting supernate, respectively adding 2 mu M hairpin DNA1 and hairpin DNA2 into the supernate, and placing the supernate in a thermostat-controlled oscillator at 37 ℃ for 40min to perform hybridization chain reaction; finally, 2.0mM rhodamine 6G is added and the reaction is carried out for 1h at room temperature. Electrochemiluminescence detection process: the reaction product formed in step (3) was added dropwise to the surface of the electrode prepared in step (1), and after reacting for 75min at room temperature, after washing three times with a phosphate buffer (pH 7.4), measurement of an electrochemiluminescence signal was subsequently performed. Electrochemiluminescence measurements were performed in a phosphate buffer (pH 7.4) containing 1mM ascorbic acid in a three-electrode system, including the prepared electrode as a working electrode, a platinum wire as a counter electrode, and a saturated calomel electrode as a reference electrode. All electrochemiluminescence signals are measured by a photomultiplier tube at the same high voltage. The DNA sequences used are all provided in table 1. As in FIG. 3A, with a constant cathode C3N4Electrochemiluminescence signal of spheres as internal standard, anode-changed CsPbBr3The electrochemiluminescence signal is used as an indicating signal, the internal reference analysis strategy is realized through the ratio of the two signals, the interference of the external environment can be eliminated, the accuracy of the analysis method is realized, the linear relation of the ratio and the cell concentration is shown in figure 3B, and the detection limit is 670 cells/mL.
TABLE 1 nucleotide sequence of the DNA used.
Example 2:
the invention provides a biological detection method for detecting the concentration of HeLa cells over-expressing carcinoembryonic antigen.
C3N4Nanosphere loaded all-inorganicPerovskite CsPbBr3The method for synthesizing the composite material comprises the following steps: 0.2035g of Cs are taken2CO35mL of octadecene and 0.5mL of oleic acid are respectively added into a three-neck flask and are placed in a vacuum drying oven at 100 ℃ for 1 h; then in N2The atmosphere was heated to 150 ℃ until the solid was completely dissolved to form a precursor solution. 0.138g of PbBr was taken25mg of hollow mesoporous C3N4Respectively adding the nanospheres (figure 1A) and 5mL of octadecene into a three-neck flask, and placing the three-neck flask in a vacuum drying oven at 100 ℃ for 1 h; followed by 1.0mL of oleylamine and 1.0mL of oleic acid, previously dried at 100 ℃ under vacuum, and then added under N2Heating to 120 deg.C in atmosphere, maintaining for 40min, and further heating to 150 deg.C to form high temperature reaction solution. And (3) quickly injecting 1.0mL of precursor solution into the high-temperature reaction solution, reacting for 3s, and then placing in an ice water bath for cooling to terminate the reaction. The reaction crude product was centrifuged at 8000rpm for 10min, the centrifuged solid was collected and washed with ethyl acetate and toluene once, respectively, and the finally collected solid was redispersed in toluene and stored for further use.
Detection of HeLa cell concentration overexpressing carcinoembryonic antigen: mix 10 uLC3N4Nanosphere loaded all-inorganic perovskite CsPbBr3The composite material is dripped on the surface of a glassy carbon electrode; after natural drying, 10 mu L of capture DNA is dripped; after 1h, dropwise adding 10 mu L of mercaptohexanol to seal the residual active sites on the surface of the material; finally, the mixture is washed clean by distilled water and is washed in N2Drying in the atmosphere. Equimolar mixing connecting DNA with carboxyl at the 5' end and functional DNA, and placing the mixture in a constant-temperature oscillator at 37 ℃ for reaction for 4 hours; then adding the carcinoembryonic antibody, 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide (EDC) and N-hydroxysuccinimide (NHS) to react for 5 hours in a constant temperature shaking instrument at 37 ℃; finally, the probe was purified by centrifugation multiple times using a 3KD ultrafiltration tube. Incubating HeLa cells with 0.5 μ M probe at 37 ℃ for 1h, centrifuging at 1000rpm for 5min, and washing the collected centrifugation precipitate with phosphate buffer (pH 7.4); after three repetitions, the final centrifugation pellet was dispersed in phosphate buffer (pH 7.4); then, the restriction enzyme DraI was added to specifically cleave the DNA probe on the cell surface in a thermostat (37 ℃) for 40min, and the cell surface was centrifugedRespectively adding 2 mu M of hairpin DNA1 and the hairpin DNA2 into the supernatant, and placing the mixture in a constant-temperature oscillator at 37 ℃ for 40min to perform hybridization chain reaction; finally, 2.0mM rhodamine 6G is added and the reaction is carried out for 1h at room temperature. Electrochemiluminescence detection process: the reaction product formed in step (3) was added dropwise to the surface of the electrode prepared in step (1), and after reacting for 75min at room temperature, after washing three times with a phosphate buffer (pH 7.4), measurement of an electrochemiluminescence signal was subsequently performed. Electrochemiluminescence measurements were performed in a phosphate buffer (pH 7.4) containing 1mM ascorbic acid in a three-electrode system, including the prepared electrode as a working electrode, a platinum wire as a counter electrode, and a saturated calomel electrode as a reference electrode. All electrochemiluminescence signals are measured by a photomultiplier tube at the same high voltage. With constant C of the cathode3N4Electrochemiluminescence signal of spheres as internal standard, anode-changed CsPbBr3The electrochemiluminescence signal is used as an indicating signal, an internal reference analysis strategy is realized through the ratio of the two signals, the interference of the external environment can be eliminated, and the accuracy of the analysis method is realized.
It should be noted that the above-mentioned embodiments are only preferred embodiments of the present invention, and are not intended to limit the scope of the present invention, and all equivalent substitutions or substitutions made on the above-mentioned embodiments are included in the scope of the present invention.
Sequence listing
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<141> 2019-12-11
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Claims (10)
1. Loaded all-inorganic perovskite CsPbBr3C of (A)3N4The synthesis method of the nanosphere composite material is characterized by comprising the following steps of:
(1) 0.2035g of Cs are taken2CO3Mixing 5-10 mL of octadecene and 0.5-0.625 mL of oleic acid, and then drying in vacuum; then heating the mixture in an inert gas atmosphere until the solid is completely dissolved to form a precursor solution;
(2) 0.138g of PbBr was taken21-5 mg of hollow mesoporous C3N4Mixing the nanospheres with 5-10 mL of octadecene, and then drying in vacuum; adding oleylamine and oleic acid under inert gasHeating to 120-150 ℃ in the atmosphere to form high-temperature reaction liquid;
(3) adding 1.0mL of the precursor solution prepared in the step (1) into the high-temperature reaction solution in the step (2), wherein the volume ratio of the precursor solution to the high-temperature reaction solution is as follows: 1: 5-10; and after reacting for 3-5 s, placing the mixture in an ice water bath for cooling to terminate the reaction, and obtaining a crude product.
2. The synthesis process of claim 1, wherein the post-treatment of the crude product obtained in step (3) is centrifugation, and the centrifuged solid is collected, washed, dispersed in toluene and stored for further use.
3. The synthesis method according to claim 1, wherein the step (1) is heating to 150 ℃ in an inert gas atmosphere until the solid is completely dissolved to form the precursor solution.
4. The synthesis method according to claim 1, wherein the vacuum drying conditions in the step (1) and the step (2) are as follows: placing in a vacuum drying oven at 100 deg.C for 1 h; inert gas atmosphere of N2And (4) atmosphere.
5. The synthesis method of claim 1, wherein in the step (2), the oleylamine and the oleic acid are dried in a vacuum oven at 100 ℃ for 1 hour in advance.
6. The synthesis method according to claim 1, wherein in the step (2), the reaction solution is heated to 120 ℃ for 40min in an inert gas atmosphere, and then the heating is continued to 150 ℃ to form a high-temperature reaction solution.
7. The supported all-inorganic perovskite CsPbBr synthesized by the synthesis method according to claim 13C of (A)3N4The application of the nanosphere composite material in detecting the number of cells is characterized by comprising the following steps:
a) the preparation process of the electrode comprises the following steps: using the loaded all-inorganic perovskite CsPbBr3C of (A)3N4The glass-carbon electrode is sequentially modified by the nanosphere composite material and the capture DNA;
b) the preparation process of the probe comprises the following steps: equimolar mixing of the connecting DNA and the functional DNA, and placing the mixture in a constant-temperature oscillator at 37 ℃ for reaction; then adding a target recognition molecule, 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide and N-hydroxysuccinimide to react in a constant temperature oscillator at 37 ℃; preparing a probe by centrifugal purification;
c) evaluation process of cell surface receptors: incubating the probe prepared in the step b) and the cells in an incubator at 37 ℃, centrifuging, collecting precipitates, washing, and dispersing in a phosphate buffer solution with the pH value of 7.4; then adding restriction enzyme DraI to specifically cut DNA probes on the cell surface in a thermostat at 37 ℃ for 40min, centrifuging, collecting supernate, adding hairpin DNA1 and hairpin DNA2 respectively, and placing in a thermostat at 37 ℃ to perform hybridization chain reaction; finally adding rhodamine 6G, and reacting at room temperature;
d) electrochemiluminescence detection process: dropwise adding the reaction product formed in the step c) to the surface of the electrode prepared in the step a), reacting at room temperature, and measuring an electrochemiluminescence signal; with constant C of the cathode3N4Electrochemiluminescence signal of spheres as internal standard, anode-changed CsPbBr3The electrochemiluminescence signal of (a) as an indicator signal, an analysis strategy of internal reference is realized by the ratio of the two signals, and the ratio has a good linear relation with the cell concentration;
the targeting molecule in step b) specifically targets the cell surface receptor of step c);
the nucleotide sequence of the connecting DNA is as follows: NH (NH)2-C6-TTT TTA AAA A；
The nucleotide sequence of the functional DNA is as follows: CCA AAC CGA AAG AAC AAT GGA CCC CTC TCC TCT CCT CTC CTC TCC TCT CCT CTC TTT TTA AAA A, respectively;
the nucleotide sequence of the hairpin DNA1 is as follows: GGG TCC ATT GTT CTT TCG GTT TGG GTA GAG CCA AAC CGA AAG AAC AAT, respectively;
the nucleotide sequence of the hairpin DNA2 is as follows: CCA AAC CGA AAG AAC AAT GGA CCC ATT GTT CTT TCG GTT TGG CTC TAC, respectively;
the nucleotide sequence of the capture DNA is as follows: SH-C6-AAA AAA AAA AGA GAG GAG AGG AGA GGA GAG GAG AGG AGA G。
8. The use of claim 7, wherein in step b), the functional DNA portion comprises an enzyme cleavage site fragment, a hybridization recognition fragment, and a priming fragment for hybridization chain reaction.
9. The use according to claim 7, wherein the electrochemiluminescence measurements of step d) are performed in a three-electrode system in phosphate buffered saline at pH 7.4 containing 1mM ascorbic acid, comprising the prepared electrode as working electrode, platinum wire as counter electrode and saturated calomel electrode as reference electrode.
10. The use of claim 7, wherein in step d), the electrochemiluminescence signals are measured by a photomultiplier tube under the same high pressure conditions.
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| CN112121837B (en) * | 2020-09-24 | 2022-05-17 | 江苏大学 | P-CN/CsPbBr3Heterojunction material, preparation method and application thereof |
| CN113265249B (en) * | 2021-05-17 | 2023-08-04 | 重庆医科大学 | Preparation and biosensing application of functionalized hydrophilic lead halide perovskite quantum dot |
| CN113189162B (en) * | 2021-05-19 | 2024-02-09 | 南昌大学 | Electrochemical luminescence method based on perovskite@covalent organic framework composite material |
| TWI792554B (en) * | 2021-09-10 | 2023-02-11 | 財團法人工業技術研究院 | Method for testing perovskite precursor solution |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107557004A (en) * | 2017-10-17 | 2018-01-09 | 南昌航空大学 | A kind of method for preparing stable perovskite quantum dot |
| CN108982478A (en) * | 2018-06-12 | 2018-12-11 | 华中师范大学 | Method based on carbonitride electrochemiluminescdetection detection chromium content |
| CN109370566A (en) * | 2018-11-26 | 2019-02-22 | 华南理工大学 | A kind of perovskite quantum dot stability improvement method adjusted based on ligand |
| CN109370563A (en) * | 2018-10-17 | 2019-02-22 | 广西大学 | A kind of halogen lead caesium perovskite fluorescent material and preparation method thereof |
| CN109706505A (en) * | 2019-01-26 | 2019-05-03 | 河南工业大学 | A kind of g-C3N4/CsPbI3/TiO2The preparation method of nano-tube array |
| CN110227532A (en) * | 2019-06-28 | 2019-09-13 | 中国计量大学 | A kind of preparation method of lead bromide caesium quantum dot/azotized carbon nano piece photochemical catalyst |
-
2019
- 2019-12-11 CN CN201911264265.1A patent/CN111518558B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107557004A (en) * | 2017-10-17 | 2018-01-09 | 南昌航空大学 | A kind of method for preparing stable perovskite quantum dot |
| CN108982478A (en) * | 2018-06-12 | 2018-12-11 | 华中师范大学 | Method based on carbonitride electrochemiluminescdetection detection chromium content |
| CN109370563A (en) * | 2018-10-17 | 2019-02-22 | 广西大学 | A kind of halogen lead caesium perovskite fluorescent material and preparation method thereof |
| CN109370566A (en) * | 2018-11-26 | 2019-02-22 | 华南理工大学 | A kind of perovskite quantum dot stability improvement method adjusted based on ligand |
| CN109706505A (en) * | 2019-01-26 | 2019-05-03 | 河南工业大学 | A kind of g-C3N4/CsPbI3/TiO2The preparation method of nano-tube array |
| CN110227532A (en) * | 2019-06-28 | 2019-09-13 | 中国计量大学 | A kind of preparation method of lead bromide caesium quantum dot/azotized carbon nano piece photochemical catalyst |
Non-Patent Citations (5)
| Title |
|---|
| All-inorganic perovskite/graphitic carbon nitride composites for CO2 photoreduction into C1 compounds under low concentrations of CO2;Siqi You et al.;《Dalton Trans.》;20190822;第48卷;第14115-14121页 * |
| Amino-Assisted Anchoring of CsPbBr3 Perovskite Quantum Dots on Porous g-C3N4 for Enhanced Photocatalytic CO2 Reduction;Man Ou et al.;《Angew. Chem. Int. Ed.》;20180917;第57卷;第13570-13574页 * |
| Fabricating CsPbX3/CN heterostructures with enhanced photocatalytic activity for penicillins 6-APA degradation;Yanyan Zhao et al.;《Chemical Engineering Journal》;20190903;第381卷;第122692（1-11）页 * |
| Improving Photovoltaic Performance Using Perovskite/Surface-Modified Graphitic Carbon Nitride Heterojunction;Zhen Li et al.;《Sol. RRL》;20191203;第4卷;第1900413（1-8）页 * |
| In situ growth of ultrasmall cesium lead bromine quantum dots in a mesoporous silica matrix and their application in flexible light-emitting diodes;Peng Chen et al.;《Nanoscale》;20190820;第11卷;第16499-16507页 * |
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