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Fluorescence imaging method for intracellular glutathione

A technique of glutathione and fluorescence imaging, applied in fluorescence/phosphorescence, material analysis through optical means, material excitation analysis, etc. simple effect

Active Publication Date: 2017-07-07
QINGDAO UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The use of gold nanocages as nanocarriers and the use of biorecognition molecules containing disulfide bonds to construct fluorescent sensors based on controllable release for the detection of GSH and fluorescence imaging technology have not been reported in the literature.

Method used

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  • Fluorescence imaging method for intracellular glutathione

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Effect test

Embodiment 1

[0020] A method for preparing the GSH fluorescent sensor based on the controllable release technology proposed by the present invention, comprising the steps of:

[0021] (1) Add 200 μL of polydiallyldimethylammonium chloride solution (11.664 mg / mL) into 400 μL of gold nanocage solution, shake at a constant temperature of 37°C for 12 hours, magnetically separate, and wash;

[0022] (2) Add 100 μL rhodamine B solution (1.0×10 -5 mol / L), 10 μL of GSH recognition probe solution (1.0×10 -5 M), shake at 37°C for 12h, magnetically separate, and wash to prepare the GSH fluorescence sensor;

[0023]Wherein, the GSH recognition probe is a nucleotide molecule containing a disulfide bond, and its base sequence is 5'-ACG AGTCA S-S TTT TGT TTG TTC CCC CCT TCT TTC TTA-3'; the gold nanocage Obtained according to literature methods (G.D.Moon, S.W.Choi, X.Cai, W.Y.Li, E.C.Cho, U.Jeong, L.V.Wang and Y.N.Xia.J.Am.Chem.Soc.2011, 133, 4762–4765).

Embodiment 2

[0025] A method for fluorescence imaging of intracellular GSH based on the controllable release technology proposed by the present invention, comprising the steps of:

[0026] (1) Centrifuge the Hela cell suspension at 3000 rpm for 5 min, and remove the supernatant;

[0027] (2) Add the PBS solution of the prepared fluorescent sensor, incubate at a constant temperature of 37°C for 20 minutes, and then perform the laser confocal scanning imaging experiment;

[0028] (3) Place the incubated cells on the stage of a confocal microscope, excitation wavelength: 559nm; reception wavelength: 580-630nm.

[0029] figure 1 Shown are fluorescence imaging photographs of GSH in Hela cells.

[0030] The invention combines molecular recognition technology, controllable release technology and nanometer technology, through specific molecular recognition reaction, can realize highly sensitive and highly selective fluorescent imaging of intracellular non-protein thiol GSH. Specifically, a holl...

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Abstract

The invention provides a fluorescence imaging method for intracellular glutathione. In the method, a fluorescence sensor based on a controlled release technology is applied to fluorescence imaging of the intracellular glutathione. The adopted fluorescence sensor comprises a gold nanometer cage, a fluorescent dye and a glutathione recognition probe, and has the advantages of simple structure, skillful design, stable performance, high cell membrane permeability, high intracellular release controllability, high glutathione selectivity, high intracellular dispersity and the like. By adopting the fluorescence sensor, high-sensitivity and high-definition fluorescence imaging of the intracellular glutathione can be realized conveniently and rapidly, and the advantages of short response time, easiness in direct observation, convenience in real-time monitoring and the like are achieved. Based on the controlled release technology, a novel technology and a novel method are provided for targeted therapy, drug delivery, cell imaging and real-time monitoring of tumor cells.

Description

technical field [0001] The invention relates to a fluorescent imaging method of glutathione (GSH) based on controllable release technology, in particular to a glutathione (GSH) in organisms, cells, and tissues that can be applied in the fields of medical care, biomedicine, and life sciences. Fluorescence Imaging Method for Glycopeptides. Background technique [0002] Glutathione (GSH) is a tripeptide compound composed of glutamic acid, cysteine, and glycine. The sulfhydryl group in cysteine ​​residues makes it an important biologically active substance. GSH widely exists in animal and plant cells, and is the most abundant non-protein thiol in cells. The content of GSH in various tissues and cells of the human body and the relationship with various diseases have attracted much attention, and have been a hot spot of extensive research so far. GSH participates in a variety of biochemical reactions in organisms, and has very important physiological functions, such as the trans...

Claims

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Application Information

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IPC IPC(8): G01N21/64
CPCG01N21/6486
Inventor 王卫董颖杰丁彩凤万均罗细亮
Owner QINGDAO UNIV OF SCI & TECH
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