Fluorescence imaging method for intracellular glutathione
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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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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.
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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 Helacell 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 laserconfocal scanning imaging experiment;
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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-proteinthiol 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...
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