CN116376038A - Preparation method of nano metal organic complex for cell imaging and copper ion detection - Google Patents
Preparation method of nano metal organic complex for cell imaging and copper ion detection Download PDFInfo
- Publication number
- CN116376038A CN116376038A CN202310093210.9A CN202310093210A CN116376038A CN 116376038 A CN116376038 A CN 116376038A CN 202310093210 A CN202310093210 A CN 202310093210A CN 116376038 A CN116376038 A CN 116376038A
- Authority
- CN
- China
- Prior art keywords
- mocps
- hdbb
- reaction kettle
- copper ions
- cell imaging
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- JPVYNHNXODAKFH-UHFFFAOYSA-N Cu2+ Chemical compound [Cu+2] JPVYNHNXODAKFH-UHFFFAOYSA-N 0.000 title claims abstract description 37
- 229910001431 copper ion Inorganic materials 0.000 title claims abstract description 37
- 238000003384 imaging method Methods 0.000 title claims abstract description 19
- 238000001514 detection method Methods 0.000 title claims abstract description 16
- 229910052751 metal Inorganic materials 0.000 title claims abstract description 9
- 239000002184 metal Substances 0.000 title claims abstract description 9
- 238000002360 preparation method Methods 0.000 title claims description 5
- 239000000463 material Substances 0.000 claims abstract description 28
- 238000000034 method Methods 0.000 claims abstract description 12
- LHPPDQUVECZQSW-UHFFFAOYSA-N 2-(benzotriazol-2-yl)-4,6-ditert-butylphenol Chemical compound CC(C)(C)C1=CC(C(C)(C)C)=CC(N2N=C3C=CC=CC3=N2)=C1O LHPPDQUVECZQSW-UHFFFAOYSA-N 0.000 claims abstract description 9
- 239000000523 sample Substances 0.000 claims abstract description 9
- 230000000694 effects Effects 0.000 claims abstract description 6
- 150000002500 ions Chemical class 0.000 claims abstract description 4
- 230000008569 process Effects 0.000 claims abstract description 4
- 238000003786 synthesis reaction Methods 0.000 claims abstract 3
- 230000015572 biosynthetic process Effects 0.000 claims abstract 2
- 238000006243 chemical reaction Methods 0.000 claims description 30
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 25
- 238000001291 vacuum drying Methods 0.000 claims description 9
- 238000001035 drying Methods 0.000 claims description 7
- 239000000243 solution Substances 0.000 claims description 7
- 238000009210 therapy by ultrasound Methods 0.000 claims description 7
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 6
- -1 polytetrafluoroethylene Polymers 0.000 claims description 6
- 230000003213 activating effect Effects 0.000 claims description 5
- 238000010438 heat treatment Methods 0.000 claims description 5
- 239000011259 mixed solution Substances 0.000 claims description 5
- 229920001343 polytetrafluoroethylene Polymers 0.000 claims description 5
- 239000004810 polytetrafluoroethylene Substances 0.000 claims description 5
- 239000000843 powder Substances 0.000 claims description 5
- 239000007787 solid Substances 0.000 claims description 5
- DUNKXUFBGCUVQW-UHFFFAOYSA-J zirconium tetrachloride Chemical compound Cl[Zr](Cl)(Cl)Cl DUNKXUFBGCUVQW-UHFFFAOYSA-J 0.000 claims description 5
- 238000000799 fluorescence microscopy Methods 0.000 claims description 4
- 239000003446 ligand Substances 0.000 claims description 4
- 238000002474 experimental method Methods 0.000 claims description 3
- 238000011895 specific detection Methods 0.000 claims description 3
- 238000004020 luminiscence type Methods 0.000 claims description 2
- 239000007864 aqueous solution Substances 0.000 claims 1
- 238000005485 electric heating Methods 0.000 claims 1
- 239000012530 fluid Substances 0.000 claims 1
- 238000012921 fluorescence analysis Methods 0.000 claims 1
- 230000006698 induction Effects 0.000 claims 1
- 238000006116 polymerization reaction Methods 0.000 claims 1
- 229910052726 zirconium Inorganic materials 0.000 claims 1
- 210000004027 cell Anatomy 0.000 abstract description 39
- 239000007850 fluorescent dye Substances 0.000 abstract description 9
- 210000000170 cell membrane Anatomy 0.000 abstract description 2
- 239000013110 organic ligand Substances 0.000 abstract description 2
- 230000035699 permeability Effects 0.000 abstract description 2
- 238000010859 live-cell imaging Methods 0.000 abstract 1
- GBNDTYKAOXLLID-UHFFFAOYSA-N zirconium(4+) ion Chemical compound [Zr+4] GBNDTYKAOXLLID-UHFFFAOYSA-N 0.000 abstract 1
- 239000000725 suspension Substances 0.000 description 5
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 4
- 229910052802 copper Inorganic materials 0.000 description 4
- 239000010949 copper Substances 0.000 description 4
- 239000012535 impurity Substances 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 239000002086 nanomaterial Substances 0.000 description 4
- 238000010791 quenching Methods 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 3
- 239000013256 coordination polymer Substances 0.000 description 3
- 229920001795 coordination polymer Polymers 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 230000005284 excitation Effects 0.000 description 3
- 230000000171 quenching effect Effects 0.000 description 3
- IJFXRHURBJZNAO-UHFFFAOYSA-N 3-hydroxybenzoic acid Chemical compound OC(=O)C1=CC=CC(O)=C1 IJFXRHURBJZNAO-UHFFFAOYSA-N 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 239000000975 dye Substances 0.000 description 2
- 238000000295 emission spectrum Methods 0.000 description 2
- 229910001385 heavy metal Inorganic materials 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 229910021645 metal ion Inorganic materials 0.000 description 2
- 239000012621 metal-organic framework Substances 0.000 description 2
- VNWKTOKETHGBQD-UHFFFAOYSA-N methane Chemical compound C VNWKTOKETHGBQD-UHFFFAOYSA-N 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 231100000419 toxicity Toxicity 0.000 description 2
- 230000001988 toxicity Effects 0.000 description 2
- 229910052723 transition metal Inorganic materials 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 239000011701 zinc Substances 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 241001338644 Retinia Species 0.000 description 1
- 108010087230 Sincalide Proteins 0.000 description 1
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 1
- 125000004429 atom Chemical group 0.000 description 1
- 238000010609 cell counting kit-8 assay Methods 0.000 description 1
- 230000011712 cell development Effects 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 231100000263 cytotoxicity test Toxicity 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000001917 fluorescence detection Methods 0.000 description 1
- 230000005283 ground state Effects 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 238000012417 linear regression Methods 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 125000004433 nitrogen atom Chemical group N* 0.000 description 1
- 229910000510 noble metal Inorganic materials 0.000 description 1
- 231100000956 nontoxicity Toxicity 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 230000010355 oscillation Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- IZTQOLKUZKXIRV-YRVFCXMDSA-N sincalide Chemical compound C([C@@H](C(=O)N[C@@H](CCSC)C(=O)NCC(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(N)=O)NC(=O)[C@@H](N)CC(O)=O)C1=CC=C(OS(O)(=O)=O)C=C1 IZTQOLKUZKXIRV-YRVFCXMDSA-N 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 230000005062 synaptic transmission Effects 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 238000012800 visualization Methods 0.000 description 1
- 229910052725 zinc Inorganic materials 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
- C08G83/00—Macromolecular compounds not provided for in groups C08G2/00 - C08G81/00
- C08G83/008—Supramolecular polymers
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N21/6456—Spatial resolved fluorescence measurements; Imaging
- G01N21/6458—Fluorescence microscopy
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
- G01N2021/6432—Quenching
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Physics & Mathematics (AREA)
- Immunology (AREA)
- Biochemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Pathology (AREA)
- General Physics & Mathematics (AREA)
- General Health & Medical Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- Polymers & Plastics (AREA)
- Optics & Photonics (AREA)
- Organic Chemistry (AREA)
- Medicinal Chemistry (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
A method for preparing nano-metal organic complex for cell imaging and copper ion detection mainly utilizes HDBB as an organic ligand and zirconium ion as a central ion to synthesize Metal Organic Complex (MOCPs). The prepared material has the advantages of high fluorescence intensity, good biocompatibility, strong cell membrane permeability and good cell imaging effect; the probe can be used for identifying and detecting copper ions with high selectivity, and the metal copper ions can be identified and detected in a living cell environment by imaging through a laser confocal microscope. The MOCPs material can be obtained through chemical synthesis, the synthesis process is simple and feasible, the popularization is easy, the fluorescent probe has high specificity, and the MOCPs material can be used for live cell imaging and real-time determination of copper ions in live cells and has wide application prospect.
Description
Technical Field
The invention relates to a method for synthesizing Zr-HDBB coordination polymer (MOCPs) by taking AIE molecules (4, 4' - (hydro-1, 2-dialkylidene) bis (3-hydroxybenzoic acid), HDBB) as an organic ligand and taking Zr with good biocompatibility as a central metal ion, belonging to the technical field of new material preparation.
Background
Living cell imaging technology is an important research tool in the field of life sciences, and can explain a variety of life and physiological phenomena in living cells. Some heavy metals and transition metal elements have very important physiological roles and functions because of being involved in the processes of cell growth and development, gene transcription, neurotransmission and the like, but when the content exceeds the corresponding threshold value, the heavy metals and the transition metal elements can cause obvious damage to the growth and development, physiological metabolism and even morphological structures of organisms [ Tsvetkov, P.; coy, s; petrova, b.; dreisppon, m.; verma, a., et al Science,2022, 375, 1254-1261; wang, g; biswas, a.k.; ma, w; kandpal, m.; coker, C., et al, nat. Med.,2018, 24, 770-781 ]. One of the most necessary methods to study the effect of specific ions on organisms is to use fluorescent probes for the fluorescence imaging detection of ions within living cells. The existing fluorescent probes for living cell imaging analysis have the defects of short fluorescence lifetime, easiness in quenching and poor biocompatibility and stability. On the other hand, copper is used as an important trace element in organisms, the content of the copper in cells is inferior to that of iron and zinc, copper ions are an important existence form of the copper element, and play an important role in the physiological and biochemical metabolism of organisms and the metabolic activity process of various enzymes, so that the development of a rapid, simple, sensitive and high-selectivity copper ion sensing detection method has important significance [ Barber, R.G.; grenier, z.a.; burkhead, j.l., biomedicines, 2021,9, 316 ]. Currently, some fluorescent probes based on carbon quantum dots (CDs), metal Organic Frameworks (MOFs), noble metal nanomaterials have been used for copper ion detection. However, in view of the specificity of living cells, some reported fluorescent probes have failed to meet the requirements for biosafety. In view of this, we have developed Metal Organic Coordination Polymers (MOCPs) with good biocompatibility. The metal organic coordination polymer has the characteristics of high fluorescence intensity, strong light stability, low biotoxicity and the like, so that cell imaging and fluorescence detection of copper ions are realized.
Disclosure of Invention
Aiming at the problems of short fluorescence life, easy quenching, poor biocompatibility and poor stability of the current fluorescent probes for living cell imaging analysis and copper ion detection, the invention aims to design and prepare MOCPs materials with good biocompatibility for cell visualization, real-time dynamic imaging and high-selectivity detection of copper ions in cells. The fluorescent nano material with high fluorescence intensity and stable optical signal in the cell environment can be prepared by the preparation method. Compared with the traditional organic dye, the fluorescent nano material has better specificity and light stability, can be efficiently absorbed by cells, has low toxicity and good biocompatibility, and can be applied to cell imaging; o atoms and N atoms in the MOCPs material are good electron donors, can show the capability of coordinating copper ions, form stable non-fluorescent ground state complexes, quench the original fluorescence of the probe, have high sensitivity and selectivity to the copper ions, and can be used for rapidly detecting the copper ions.
The technical scheme of the invention is as follows:
33.2 mg (0.1 mmol) of HDBB and 64.85 mg (0.2 mmol) of zirconium chloride were weighed and dissolved in 15 mL of N, N-dimethylformamide.
After ultrasonic treatment for 10 minutes, the obtained mixed solution is transferred into a reaction kettle with 50 mL polytetrafluoroethylene lining, the reaction kettle is repeatedly screwed up, and then the reaction kettle is placed in an electrothermal constant-temperature drying oven for heating reaction at 120 ℃ for 24 h.
After the reaction is finished and the reaction kettle is naturally cooled to room temperature, a centrifuge (8000 rpm, 8 min) is used for removing larger impurities, and then supernatant is extracted for later use. Washing with 20-30 mL DMF at least 3 times, washing with 20-30 mL absolute ethanol at least 3 times, and drying at 60 ℃. Vacuum drying and activating for 10-12 hours at 60 ℃ in a vacuum drying oven to obtain light yellow solid powder, namely Zr-HDBB material for standby.
And (3) culturing the MOCPs material solution and the cells in a constant temperature incubator, observing the internal structure of the cells on a laser confocal microscope under the condition of luminescence of fluorescent nano materials, and taking cell imaging pictures.
When copper ions exist, the copper ions react with the probe, causing significant changes in the fluorescence intensity of the MOCPs. The specific detection of copper ions was achieved by fluorescence intensity measurement on a molecular fluorescence photometer (excitation wavelength 365 nm).
Compared with the prior art, the invention has the following advantages:
the detection sensitivity is high, and the selectivity is good;
the dosage of the detection reagent is small, and the detection method is simple and quick to operate;
the probe has good cell membrane permeability, low cytotoxicity and good cell imaging effect;
the probe has good response to copper ions, but can be used for imaging and detecting copper ions in cells.
Description of the embodiments
Example 1
33.2 mg (0.1 mmol) of the HDBB ligand and 64.85 mg (0.2 mmol) of zirconium tetrachloride were weighed out and dissolved in 15 mL of N, N-dimethylformamide. After ultrasonic treatment for 10 minutes, the obtained mixed solution is transferred into a reaction kettle with 50 mL polytetrafluoroethylene lining, the reaction kettle is repeatedly screwed up, and then the reaction kettle is placed in an electrothermal constant-temperature drying oven to react under heating at 120 ℃ for 48 h. After the reaction is finished and the reaction kettle is naturally cooled to room temperature, removing larger impurities by using a centrifugal machine (8000 rpm, centrifuging for 10 min), and extracting supernatant for later use. Washed 3 times with 30 mL DMF, 1 time with 10 mL secondary water, and finally 3 times with 30 mL absolute ethanol, and then dried at 60 ℃. Vacuum drying and activating for 12 hours at 60 ℃ in a vacuum drying oven to obtain light yellow solid powder, namely Zr-HDBB material for standby.
The prepared MOCPs material is prepared into a suspension liquid of 1.0 mg/mL, ice bath is carried out for 30 min, and the fluorescence excitation wavelength is 365nm, and the emission spectrum is measured by using a fluorescence spectrophotometer. Respectively adding copper ion to-be-detected solutions with different concentrations into the fluorescent probe solution, carrying out slight oscillation reaction on the solutions for 5 min at room temperature, measuring an emission spectrum by using a fluorescence spectrophotometer under the same condition by taking 365nm as fluorescence excitation wavelength, and verifying the detection effect; recording the change value F of fluorescence intensity 0 /F（F 0 The fluorescence intensity of MOCPs at a copper ion concentration of 0. Mu.M, and F is the fluorescence intensity of MOCPs after copper ion addition). Reduce the concentration of copper ions, F 0 The value of/F was also correspondingly reduced, indicating that the fluorescence intensity of the MOCPs was related to the copper ion concentration. In the range of 0-200 mu M of copper ion concentration, the change of fluorescence intensity and the copper ion concentration show good linear relation, the quantitative detection of copper ions can be realized, and a fitted linear regression equationY=0.0579 x+1.123. Interference tests showed that other various metal ions (Al 3+ ，Ca 2+ ，Co 2+ ，Fe 3+ ，K + ，Mg 2+ ，Na + ，Ni 2+ ，Zn 2+ ) The existence of (1) does not interfere with the identification of copper ions by MOCPs materials; the fluorescent probe can realize the specific detection of copper ions.
Examples
33.2 mg (0.1 mmol) of the HDBB ligand and 64.85 mg (0.2 mmol) of zirconium tetrachloride were weighed out and dissolved in 15 mL of N, N-dimethylformamide. After ultrasonic treatment for 10 minutes, the obtained mixed solution is transferred into a reaction kettle with 50 mL polytetrafluoroethylene lining, the reaction kettle is repeatedly screwed up, and then the reaction kettle is placed in an electrothermal constant-temperature drying oven to react under heating at 120 ℃ for 36 h. After the reaction is finished and the reaction kettle is naturally cooled to room temperature, a centrifuge (8000 rpm, 15 min) is used for removing larger impurities, and then supernatant is extracted for later use. Washed 3 times with 30 mL DMF, 3 times with 30 mL absolute ethanol, and then dried at 60 ℃. Vacuum drying and activating for 10 hours at 60 ℃ in a vacuum drying oven to obtain light yellow solid powder, namely Zr-HDBB material for standby.
Firstly, determining the toxicity influence of MOCPs material on cells, and using A549 cells to carry out cytotoxicity test by CCK-8 method, wherein the cell survival rate is above 85%, which shows that the synthesized MOCPs material has almost no toxicity and good biocompatibility, and can be used for cell imaging.
The prepared MOCPs material was prepared into a suspension of 1.0. 1.0 mg/mL, ice-bath sonicated for 30 min, and 1.0. 1.0 mg/mL of MOCPs material was diluted to 50. Mu.g/mL with cell-specific medium. The diluted probe solution and A549 cells are co-cultured for 1 h, and observed under a laser confocal microscope, MOCPs can enter the interiors of the A549 cells and emit bright red fluorescence. Compared with the traditional dye, the MOCPs have better light stability, can be efficiently absorbed by cells, can successfully enter the cells and realize fluorescence imaging of the cells.
Examples
33.2 mg (0.1 mmol) of the HDBB ligand and 64.85 mg (0.2 mmol) of zirconium tetrachloride were weighed out and dissolved in 15 mL of N, N-dimethylformamide. After ultrasonic treatment for 10 minutes, the obtained mixed solution is transferred into a reaction kettle with 50 mL polytetrafluoroethylene lining, the reaction kettle is repeatedly screwed up, and then the reaction kettle is placed in an electrothermal constant-temperature drying oven for heating reaction at 120 ℃ for 24 h. After the reaction is finished and the reaction kettle is naturally cooled to room temperature, removing larger impurities by using a centrifugal machine (10000 rpm, 10 min), and extracting supernatant for later use. Washed 3 times with 30 mL DMF, 1 time with 10 mL secondary water, and finally 3 times with 30 mL absolute ethanol, and then dried at 60 ℃. Vacuum drying and activating for 10 hours at 60 ℃ in a vacuum drying oven to obtain light yellow solid powder, namely Zr-HDBB material for standby.
The prepared MOCPs material is prepared into a suspension liquid of 1.0 mg/mL, the suspension liquid is subjected to ice bath ultrasonic treatment for 30 min, and the suspension liquid of the material after ultrasonic treatment is diluted to 50 mug/mL by using a cell special culture medium. To demonstrate that the probe was able to detect copper ions in a biological system, 50 μg/mL of MOCPs material was first co-cultured with a549 cells for 3 h, then the culture broth of copper-containing material was added for continued culture for 1 h. After three washes with PBS buffer, imaging was performed using a laser confocal microscope, and significant quenching of intracellular fluorescence was observed. Throughout the experiment, the cells were still viable and visualized, indicating that the fluorescent probes were free of significant toxicity and side effects. The fluorescence imaging experiment shows that the MOCPs material can enter cells and can be used as a fluorescence probe for detecting copper ions in the cells, and has wide application prospect in the practical fields of cell imaging, in-vivo detection of copper ions and the like.
Claims (5)
1. A preparation method of Zr-HDBB Metal Organic Complex (MOCPs) nano-probes for cell imaging and copper ion detection is characterized by comprising the following steps:
(1) 33.2 mg (0.1 mmol) of HDBB and 64.85 mg (0.2 mmol) of zirconium tetrachloride were weighed and dissolved in 15 mL of N, N-dimethylformamide;
(2) After ice bath ultrasonic treatment for 10 minutes, transferring the obtained mixed solution into a reaction kettle with 50 mL polytetrafluoroethylene lining, repeatedly screwing the reaction kettle, and then placing the reaction kettle into an electric heating constant temperature drying oven to perform heating reaction for 24-48 hours at 120 ℃;
(3) After the reaction is finished and the reaction kettle is naturally cooled to room temperature, the reaction kettle is washed for a plurality of times by using DMF and ethanol and then is dried at 60 ℃. Activating and drying to obtain yellowish solid powder, namely a coordination induction luminescent material Zr-HDBB material;
(4) After incubating MOCPs solution with cells, performing a fluorescence imaging experiment by using a laser confocal microscope;
(5) MOCPs solution reacts with copper ions outside or inside cells, and the specific detection of aqueous solution samples, human tissue fluid and copper ions inside cells is realized through a fluorescence analysis method.
2. The process of claim 1, wherein the core ion of the MOCPs material is Zr 3+ The ligand is a polymerization-induced luminescent molecule HDBB.
3. The method of claim 1, wherein zirconium ions in the prepared MOCPs material are capable of being linked to HDBB molecules by metal coordination bonds to produce a coordination induced luminescence effect.
4. The method of claim 1, wherein the material is activated by vacuum drying in a vacuum oven at 60 ℃ for 10-12 hours after synthesis.
5. The method of claim 1, wherein the prepared MOCPs material is useful for cell imaging and copper ion detection.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310093210.9A CN116376038A (en) | 2023-02-10 | 2023-02-10 | Preparation method of nano metal organic complex for cell imaging and copper ion detection |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310093210.9A CN116376038A (en) | 2023-02-10 | 2023-02-10 | Preparation method of nano metal organic complex for cell imaging and copper ion detection |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN116376038A true CN116376038A (en) | 2023-07-04 |
Family
ID=86970069
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310093210.9A Pending CN116376038A (en) | 2023-02-10 | 2023-02-10 | Preparation method of nano metal organic complex for cell imaging and copper ion detection |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116376038A (en) |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1220291A (en) * | 1997-09-05 | 1999-06-23 | 三菱综合材料株式会社 | Transparent conductive film and composition for forming same |
| CN105523960A (en) * | 2016-02-25 | 2016-04-27 | 云南大学 | Synthetic method of novel pH-response double-emission-wavelength fluorescence molecular probe and application of novel pH-response double-emission-wavelength fluorescence molecular probe in bio-imaging |
| US20180200553A1 (en) * | 2016-02-26 | 2018-07-19 | Sinochem Lantian Co., Ltd | Composition comprising fluorine-containing ketone |
| JP2019015610A (en) * | 2017-07-07 | 2019-01-31 | 日本化薬株式会社 | Ultraviolet curable resin composition used in bonding of biosensor board and biosensor board using the same |
| US20200087451A1 (en) * | 2018-09-15 | 2020-03-19 | The Board Of Regents Of The University Of Texas System | Dual modality ups nanoprobes for tumor acidosis imaging |
| CN113185969A (en) * | 2021-04-29 | 2021-07-30 | 浙江大学 | Fluorescent film for real-time detection of metal ions in water body and preparation method thereof |
| US11124530B1 (en) * | 2019-10-22 | 2021-09-21 | King Fahd University Of Petroleum And Minerals | Zirconium metal-organic framework and a method of detecting copper and chromate ions |
| CN113480731A (en) * | 2021-07-22 | 2021-10-08 | 西安工业大学 | Detectable Cu2+And ClO-Preparation method of fluorescent carbon dot and hydrogel |
-
2023
- 2023-02-10 CN CN202310093210.9A patent/CN116376038A/en active Pending
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1220291A (en) * | 1997-09-05 | 1999-06-23 | 三菱综合材料株式会社 | Transparent conductive film and composition for forming same |
| CN105523960A (en) * | 2016-02-25 | 2016-04-27 | 云南大学 | Synthetic method of novel pH-response double-emission-wavelength fluorescence molecular probe and application of novel pH-response double-emission-wavelength fluorescence molecular probe in bio-imaging |
| US20180200553A1 (en) * | 2016-02-26 | 2018-07-19 | Sinochem Lantian Co., Ltd | Composition comprising fluorine-containing ketone |
| JP2019015610A (en) * | 2017-07-07 | 2019-01-31 | 日本化薬株式会社 | Ultraviolet curable resin composition used in bonding of biosensor board and biosensor board using the same |
| US20200087451A1 (en) * | 2018-09-15 | 2020-03-19 | The Board Of Regents Of The University Of Texas System | Dual modality ups nanoprobes for tumor acidosis imaging |
| US11124530B1 (en) * | 2019-10-22 | 2021-09-21 | King Fahd University Of Petroleum And Minerals | Zirconium metal-organic framework and a method of detecting copper and chromate ions |
| CN113185969A (en) * | 2021-04-29 | 2021-07-30 | 浙江大学 | Fluorescent film for real-time detection of metal ions in water body and preparation method thereof |
| CN113480731A (en) * | 2021-07-22 | 2021-10-08 | 西安工业大学 | Detectable Cu2+And ClO-Preparation method of fluorescent carbon dot and hydrogel |
Non-Patent Citations (4)
| Title |
|---|
| LU, ZX , 等: "Modulating the optical properties of the AIE fluophor confined within UiO-66\'s nanochannels for chemical sensing", NANOSCALE, vol. 8, no. 40, 16 November 2016 (2016-11-16), pages 17489 - 17495 * |
| ZHAO, JF，等: "Effects of solvent polarity on excited state behaviors for the two intramolecular proton-transfer-site 4, 4′-(hydrazine-1, 2-diylidene-bis (methanylylidene))-bis(3-hydroxybenzoic acid) compound", JOURNAL OF LUMINESCENCE, vol. 232, 5 March 2021 (2021-03-05), pages 1 - 8 * |
| 张琴: "AIE分子修饰的金属-有机框架物的荧光性质及其应用研究", 中国优秀硕士学位论文全文数据库 (工程科技Ⅰ辑), no. 01, 15 January 2020 (2020-01-15), pages 014 - 446 * |
| 林娜: "AIE分子功能化的配合物纳米粒子的合成及其应用", 中国优秀硕士学位论文全文数据库 (工程科技Ⅰ辑), no. 02, 15 February 2017 (2017-02-15), pages 014 - 876 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Qu et al. | A red fluorescent turn-on probe for hydrogen sulfide and its application in living cells | |
| CN106905310B (en) | It is a kind of to detect hypochlorous fluorescence probe and its preparation method and application | |
| Li et al. | A near-infrared fluorescent probe for Cu2+ in living cells based on coordination effect | |
| CN105712964B (en) | Preparation method and application of thiol fluorescent probe based on coumaroyl hydrazide | |
| CN112646191B (en) | Rare earth-organic framework nano fluorescent probe and preparation method and application thereof | |
| CN111285833A (en) | Detection ONOO-Ratiometric fluorescent molecular probe and preparation method and application thereof | |
| Wang et al. | A novel “turn-on” fluorogenic probe for sensing hypochlorous acid based on BODIPY | |
| Li et al. | A near-infrared fluorescent probe for monitoring tyrosinase activity | |
| Yan et al. | An iridium complex-based probe for photoluminescence lifetime imaging of human carboxylesterase 2 in living cells | |
| CN112159522B (en) | Water-soluble rhodamine-based fluorescent/colorimetric dual-mode probe and preparation method and application thereof | |
| Han et al. | A ratiometric nanoprobe consisting of up-conversion nanoparticles functionalized with cobalt oxyhydroxide for detecting and imaging ascorbic acid | |
| CN105154065B (en) | A kind of fluorescent probe of quick specific recognition hydroxyl radical free radical and its preparation method and application | |
| CN109932349B (en) | Organic silicon micromolecule fluorescent probe for detecting hypochlorous acid | |
| Li et al. | Pyridine functionalized carbon dots for specific detection of tryptophan in human serum samples and living cells | |
| CN110818646A (en) | Aggregation-induced emission-based small-molecule fluorescent probe and preparation method and application thereof | |
| CN108752373B (en) | Fluorescent probe for identifying hydrogen peroxide based on phenylboronate | |
| Zhang et al. | Based on theoretical calculations designed a novel dual-channel chemo-sensor for Mg2+ and Zn2+ detection and bioimaging applications | |
| CN110372681B (en) | Application of self-assembled nano fluorescent probe for selectively detecting human serum albumin | |
| CN110357896B (en) | Compound, preparation and application thereof in detecting divalent copper ions and strong acid pH | |
| CN111349071A (en) | For detecting ONOO-Xanthene derivative, and synthetic method and application thereof | |
| CN116376038A (en) | Preparation method of nano metal organic complex for cell imaging and copper ion detection | |
| CN112358475B (en) | Benzopyrylium-phenothiazine derivative and preparation method and application thereof | |
| CN110643044B (en) | Fluorescent probe for detecting hypochlorous acid and preparation method and application thereof | |
| CN114636746A (en) | Detect Pb2+Carboxyl ligand induced annihilation type ratio electrochemiluminescence aptamer sensing method | |
| CN110687087A (en) | Preparation method and application of lysosome adenosine triphosphate recognition carbon dots |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |