Nano probe for copper ion fluorescence imaging in cells and preparation method for nano probe

A technology of fluorescence imaging and nanoprobes, which is applied in preparations for in vivo experiments, fluorescence/phosphorescence, chemical instruments and methods, etc., can solve the unsatisfactory chemical stability of quantum dot fluorescent probes, organic molecular probes, etc. Unstable structure, high cell lethality, etc., to avoid light damage, good shape, and avoid strong biological background fluorescence

Inactive Publication Date: 2015-05-06
SHANGHAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

They have the following disadvantages: (1) Organic molecular probes are unstable in structure, poor in photostability, and poor in water solubility, so they cannot be applied in pure aqueous environments or cell systems
(2) The chemical stability of quantum dot-based fluorescent probes is still unsatisfactory, and its biological toxicity is still controversial.
(3) The excitation light of the probe used is located in the high-energy light region such as the ultraviolet and visible light region, which is highly lethal to cells, and at the same time has strong biological autofluorescence and low signal-to-noise ratio

Method used

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  • Nano probe for copper ion fluorescence imaging in cells and preparation method for nano probe
  • Nano probe for copper ion fluorescence imaging in cells and preparation method for nano probe
  • Nano probe for copper ion fluorescence imaging in cells and preparation method for nano probe

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preparation example Construction

[0026] For the preparation method of upconversion luminescent nanocrystals with core-shell structure, please refer to the literature:

[0027] 27. Li, Z.; Zhang, Y., An efficient and user-friendly method for the synthesis of hexagonal-phase NaYF 4 :Yb, Er / Tm nanocrystals with controllable shape and upconversion fluorescence. Nanotechnology 2008, 19 (34), 345606.

Embodiment 1

[0029] This embodiment provides an example to nanoprobe CS-UCNPsmSiO 2 -The synthesis of RBH, it comprises the following steps:

[0030] (1) Upconversion nanoparticles CS-UCNPs with core-shell structure were synthesized by solvothermal method, and dispersed into cyclohexane solution to obtain upconversion nanoparticles with core-shell structure;

[0031] (2) Using hexadecyltrimethylammonium bromide as a template and surfactant, the upconverting nanoparticles with core-shell structure were transferred from cyclohexane solution to aqueous phase, and then used in an alkaline environment The hydrolysis and condensation reaction of tetraethyl orthosilicate coats the surface of nanoparticles with a layer of silicon dioxide, and then removes the surfactant by ion exchange to obtain mesoporous-coated upconversion nanoparticles with composite functions, namely the preparation of CS-UCNPsmSiO 2 ;

[0032] (3) Reacting rhodamine B derivative fluorescent dyes with triethoxysilyl isocya...

Embodiment 2

[0052] The present embodiment provides the spectrograms of nanoprobes and different metal ions, which includes the following steps:

[0053] (1) Concentrated solutions (0.1M) of various metal ions are prepared with deionized water;

[0054] (2) Preparation of nanoprobe CS-UCNPsmSiO 2 -RBH, then diluted to the desired concentration with absolute ethanol solution.

[0055] (3) In the selectivity experiment, a certain volume of concentrated solutions of various metal ions was added to 2.0mL nanoprobe CS-UCNPsmSiO 2 -RBH solution, stirred at room temperature for 1 min, then carried out spectral test, and observed its color change. The result is as figure 2 shown.

[0056] (4) In the titration experiment, pipette 2.0mL nanoprobe CS-UCNPsmSiO 2 - RBH is placed in a 1.0cm×1.0cm quartz cuvette, and Cu is added dropwise to the secondary solution with a micro-injector 2+ solution, every time a certain volume of Cu was added 2+ After the solution was stirred at room temperature f...

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Abstract

The invention relates to a nano probe for copper ion fluorescence imaging in cells and a preparation method for the nano probe. The preparation method comprises the following steps: firstly, synthesizing up-conversion nana particles with core-shell structures, CS-UCNPs for short; then, by taking a CTAB (cetyltrimethyl ammonium bromide) as a template agent and a surfactant, coating a layer of mesoporous SiO2 on the surfaces of the CS-UCNPs to obtain mesoporous coated nano particles CS-UCNPs@mSiO2 with compounding functions; finally, performing covalent bond assembly on functional modified rhodamine organic dye RHB precursors capable of identifying metal copper ions in a singular manner and the CS-UCNPs@mSiO2 to construct an organic-inorganic hybrid nano probe CS-UCNPs@mSiO2-RBH, thereby realizing fluorescence detecting and living cell fluorescence imaging of copper ions in an aqueous solution. The preparation method disclosed by the invention has the advantages of being simple in process, convenient to operate and easy to control structure. The prepared nano fluorescence probe has the characteristics of a uniform dimension, a stable structure, low toxicity and good biocompatibility, and a potential application value in the fields of up-conversion fluorescence labeling, fluorescence detecting and the like of cells or tissues.

Description

technical field [0001] The invention relates to a nanometer probe for intracellular copper ion fluorescence imaging and a preparation method thereof. Background technique [0002] Copper is an essential trace element in the human body and plays an important role in the metabolic process of the human body. Copper deficiency will reduce the pigment oxidase and vitality in brain cells, resulting in memory loss, thinking disorder, unresponsiveness, and even unsteady gait, movement disorders, etc.; copper participates in the synthesis of various metalloenzymes in the human body, Once the copper element is deficient, the synthesis of such enzymes will be reduced, and the cardiovascular system will not be able to maintain its normal shape and function, thus giving an opportunity for the invasion of coronary heart disease. On the other hand, excessive copper intake may cause poisoning, including acute copper poisoning, hepatolenticular degeneration, and intrahepatic cholestasis in ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C09K11/85C09K11/06A61K49/00G01N21/64
Inventor 施利毅刘金亮徐艳霞孙丽宁孟宪福
Owner SHANGHAI UNIV
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