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an intracellular ag + fluorescence imaging method
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A fluorescence imaging and cell technology, applied in the field of intracellular Ag+ fluorescence imaging, to achieve the effects of stable performance, strong cell membrane permeability and simple structure
Active Publication Date: 2022-03-11
北京初美人医疗美容门诊部有限公司
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Gold nanoparticles with a hollow and porous structure are used as nanocarriers, and C-C base mismatches are used to pair Ag + Constructing nanocomposites for detection of Ag with unique biorecognition + and its fluorescence imaging technology have not been reported in the literature
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Embodiment 1
[0022] A method for preparing a nano-gold composite material based on base mismatch recognition technology proposed by the present invention, comprising the steps of:
[0023] (1) Design and synthesize Ag + Cytosine-rich nucleic acid biomolecules with a certain base length, which are recognized by base mismatches, and which can interact with Ag + A specific base mismatch recognition reaction occurs;
[0024] (2) Centrifuge the nano-gold carrier solution (400 μL) with a hollow and porous structure, remove the supernatant, add polydiallyldimethylammonium chloride solution (200 μL, 11.664 mg / mL), and after 10 h at 37 ° C Centrifuge and remove the supernatant;
[0025] (3) Add rhodamine B solution (2μL, final concentration 1.0×10 -4 mol / L), dilute to 100 μL with MOPS buffer solution with pH=7.0, add identifiable Ag after 10 h at 37 °C + Biomolecule solution (10 μL, final concentration 1.0×10 -6 mol / L), centrifuged at 37°C for 10 hours, removed the supernatant, and diluted wi...
Embodiment 2
[0028] A gold nanocomposite based on base mismatch recognition technology for intracellular Ag + The fluorescence imaging method is as follows:
[0029] (1) Take an appropriate amount of cell suspension, centrifuge, remove the supernatant, and use 100 μL for a final concentration of 1.0×10 -4 Cultivate the cells with silver nitrate solution of M, centrifuge at 37°C for 20.0min, and remove the supernatant;
[0030] (2) Add the solution prepared in Example 1, place the cells in a constant temperature water bath at 37°C to incubate the cells for 15.0 min, and then conduct laser confocal scanning imaging;
[0031] (3) Place the incubated cells on the stage of a confocal microscope, excitation wavelength: 559nm; reception wavelength: 580-630nm;
[0032] Wherein, the cells are cervical cancer cells.
[0033] figure 1 Ag in cervical cancer cells + fluorescence imaging photos. The results show that when the nano-gold composite material based on the base mismatch recognition tech...
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Abstract
The present invention proposes a kind of Ag in the cell + Fluorescent imaging method, which can be combined with Ag by designing and synthesizing + Biomolecules recognized by base mismatches are assembled onto the surface of nano-gold carriers with hollow and porous structure characteristics, and nano-gold composite materials with "pore caps" are constructed. When it enters the cell, the intracellular Ag + Due to the interaction with the biomolecules on the surface of the nanocarriers, the biomolecules are detached from the surface of the nanocarriers, the "pore cap" is opened, and the dye molecules in the nanocarriers are released, realizing the detection of Ag in the cells. + fluorescence imaging. The nano-gold composite material based on the base mismatch recognition technology adopted in the present invention has the advantages of simple structure, ingenious design, stable performance, strong cell membrane penetration ability, strong controllability of intracellular release, and strong anti-Ag + With many advantages such as good selectivity, short response time, and easy real-time monitoring, it can be convenient and fast to realize the detection of intracellular Ag + Highly sensitive and selective fluorescence imaging.
Description
technical field [0001] The present invention relates to a kind of intracellular Ag + Fluorescent imaging method, specifically related to a base mismatch recognition technology based on intracellular Ag + fluorescence imaging method. Background technique [0002] Silver ions as heavy metal ions (Ag + ) and mercury ions (Hg 2+ ) is similar, it is a metal ion with high toxicity, and it is also a widely distributed environmental pollutant. Even at low concentrations, it can cause serious and permanent toxicity to the environment and humans. More seriously, through polluted water sources, Ag + It can enter the human food chain after accumulating in agricultural and aquatic products. If prolonged exposure to Ag + The environment in which it exists may lead to the slow occurrence of human degenerative diseases in the body and nervous system. Therefore, the establishment of efficient, sensitive and economical Ag + The detection method is of great significance in the fields ...
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