Method for simultaneously detecting hypochlorite ions and zinc ions in cells based on up-conversion fluorescence probe

A technology of hypochlorite and fluorescent probes, applied in chemical instruments and methods, fluorescence/phosphorescence, luminescent materials, etc.

Active Publication Date: 2021-08-27
FUJIAN INST OF RES ON THE STRUCTURE OF MATTER CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to solve the problem that the up-conversion fluorescent probes in the prior art cannot realize accurate quantitative detection of intracellular hypochlorite ions and zinc ions at the same time, the present invention provides an up-conversion fluorescent probe based on double excitation and double emission to simultaneously detect hypochlorite ions and zinc ions in cells. A method for chlorate ions and zinc ions, the method is to realize multiple accurate detection of hypochlorite ions and zinc ions in cells through double-excitation and double-emission modes

Method used

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  • Method for simultaneously detecting hypochlorite ions and zinc ions in cells based on up-conversion fluorescence probe
  • Method for simultaneously detecting hypochlorite ions and zinc ions in cells based on up-conversion fluorescence probe
  • Method for simultaneously detecting hypochlorite ions and zinc ions in cells based on up-conversion fluorescence probe

Examples

Experimental program
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Effect test

preparation example 1

[0106] Preparation of double excitation double emission upconversion fluorescent probe UCNP-IR845-PAPS-F127:

[0107] (1) 1mmol oil-soluble nanoparticles NaYF 4 :Yb,Er@NaYF 4 : Yb and 1 μmol of the dye IR845 were dispersed in 20 mL of solvent chloroform, mixed for one hour to obtain solution 1;

[0108] (2) Dissolve 40mg of F127 in 2mL of chloroform, and add it dropwise to solution 1 under constant stirring to obtain solution 2;

[0109] (3) The solution two is rotary evaporated to remove the solvent trichloromethane, and after the volatilization of the trichloromethane is completed, it is redispersed in the aqueous solution to obtain the solution three;

[0110] (4) 48 μmol of visible light dye molecule PAPS is evenly dispersed in the aqueous solution, and the PAPS aqueous solution is added dropwise to the solution three under constant stirring to obtain solution four, and the stirring is continued overnight;

[0111] (5) After the step (4) is completed, centrifuge and was...

Embodiment 1

[0112] Example 1 Detection of hypochlorite ions by double-excitation and double-emission up-conversion fluorescent probes:

[0113] (1) Disperse the up-converting fluorescent probe prepared in Preparation Example 1 above in water to prepare a dispersion solution with a concentration of 200 mM. , 70, 80, 90 and 100 μM target detection substance (hypochlorite ion) reacted for 30 minutes;

[0114] (2) Measure the upconversion luminescence intensity of the solution in the step (1) on the FLS980 fluorescence spectrometer;

[0115] (3) Take the concentration of the hypochlorous acid to be measured in the mixed solution as the abscissa, and the up-conversion luminescence intensity corresponding to 830nm excitation as the ordinate to draw the concentration-dependent standard curve of the hypochlorous acid, and calculate the detection limit.

[0116] Such as Figure 5 As shown, in the range of 0-50 μM, the higher the concentration of hypochlorite ions, the lower the corresponding up-...

Embodiment 2

[0117] Example 2 Utilizes double-excitation double-emission up-conversion fluorescent probe to detect zinc ions:

[0118] (1) Disperse the up-converting fluorescent probe prepared in Preparation Example 1 above in water to prepare a dispersion solution with a concentration of 200 mM. , 70 and 80 μM target detection substance (zinc ion) reacted for 30 minutes;

[0119] (2) Measure the upconversion luminescence intensity of the solution in the step (1) on the FLS980 fluorescence spectrometer;

[0120] (3) Taking the concentration of zinc ions to be measured in the mixed solution as the abscissa, and the up-conversion luminescence intensity corresponding to 980nm excitation as the ordinate, draw the concentration-dependent standard curve of the zinc ions, and calculate the detection limit.

[0121] Such as Figure 6 As shown, in the range of 0-27 μM, the higher the concentration of zinc ions, the lower the intensity of upconverted green light corresponding to the mixed solution...

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Abstract

The invention belongs to the technical field of nano biological materials, and particularly relates to a method for simultaneously detecting hypochlorite ions and zinc ions in cells based on an up-conversion fluorescence probe. According to the method, through a near-infrared dual-laser confocal microscope system, multiple quantitative detection on substances in cells can be realized by utilizing a dye-sensitized / quenched up-conversion fluorescence probe. Compared with a traditional fluorescent probe detection method, the method disclosed by the invention has the advantages that hypochlorous acid and zinc ions in cells can be accurately detected at the same time by utilizing two energy transfer processes of dye-sensitized / quenched upconversion nanoparticles, and a double-excitation double-emission-ratio probe model is provided as a set reference; the detection deviation caused by complex environment, probe distribution and instrument equipment in the cells is reduced, and a new idea is provided for simultaneously detecting various substances in the cells.

Description

technical field [0001] The invention belongs to the technical field of nano-biological materials, and in particular relates to a method for simultaneously detecting intracellular hypochlorite ions and zinc ions based on a double-excitation double-emission up-conversion fluorescent probe. Background technique [0002] Intracellular multiple detection analysis can simultaneously qualitatively or quantitatively monitor the concentration changes of various ions, chemical molecules, proteins, nucleic acids and other components in cells, and has received more and more attention in the fields of biomedicine and biochemistry. Simultaneous multiplexing of intracellular chemicals provides greater insight into cellular function and reveals biochemical processes in disease development. Compared with the traditional single analyte, the analysis speed of the multiplex detection is faster, the consumption of reagents is lower, and the labor required is less, thus greatly improving the effi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64C09K11/06
CPCG01N21/6428C09K11/06G01N2021/6432G01N2021/6419G01N2021/6421C09K2211/14
Inventor 陈学元卢珊柯建熙李幸俊李卓
Owner FUJIAN INST OF RES ON THE STRUCTURE OF MATTER CHINESE ACAD OF SCI
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