a cu + Fluorescent probe and its preparation method and application

A fluorescent probe and probe technology, applied in the fields of fluorescence imaging and biosensing, can solve the problems of low signal interference, achieve the effects of correcting environmental interference, improving accuracy, and enriching detection methods

Active Publication Date: 2020-01-07
EAST CHINA NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Realize the application of Cu in cells and living bodies + The precise detection of probes requires high selectivity and low signal interference. Therefore, the development has been able to realize Cu in cells and in vivo + The precise detection of probes is challenging

Method used

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  • a cu <sup>+</sup> Fluorescent probe and its preparation method and application
  • a cu <sup>+</sup> Fluorescent probe and its preparation method and application
  • a cu <sup>+</sup> Fluorescent probe and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0109] Example 1. Cu + Preparation of fluorescent probes

[0110] (1) Preparation of copper ion ligand: 2.12g hydroxyethyl sulfide (20mmol) and 1.52g thiourea (20mmol) were mixed and reacted in 4.3mL 48% hydrobromic acid (32mmol), and refluxed overnight under nitrogen atmosphere. The reaction was cooled to 25° C., aqueous sodium hydroxide solution (40 mmol) was slowly added, and the mixed reaction product was refluxed overnight under nitrogen atmosphere. Then the reaction was cooled to 25°C, neutralized with concentrated hydrochloric acid, then extracted with 100 mL of ethyl acetate, the organic phase was separated, washed with 100 mL of water, then dried with magnesium sulfate, and evaporated to dryness to obtain a colorless, Pungent smelling oil (1.7 g, 70% yield).

[0111] Then, 0.56 g of sodium (23.3 mmol) and 0.34 g of the above product (2.8 mmol) were added to 60 mL of absolute ethanol under a nitrogen atmosphere, and the mixed solution was heated to reflux. 0.66g of ...

Embodiment 2

[0117] Example 2. Detection of Cu in an in vitro environment +

[0118] (1) Preparation of fluorescence intensity calibration curve

[0119] Get the Cu prepared in 1mL embodiment 1 + Fluorescent probes, diluted to 2mL with phosphate buffer, added 10μL of equal concentration of Cu each time + solution, and then measure the fluorescence intensity of the solution with an excitation wavelength of 405 nm. As the added volume increases, the fluorescence intensity of the probe decreases gradually, and shows good linearity in the range of 5-80μM, the lowest detection line is 0.60μM; the linear relationship is R 2 =0.997( Figure 4 ).

[0120] (2) Preparation of fluorescence lifetime and calibration curve

[0121] Get the Cu prepared in 1mL embodiment 1 + Fluorescent probes, diluted to 2mL with phosphate buffer, added 10μL of equal concentration of Cu each time + solution, and then measure the fluorescence intensity of the solution with an excitation wavelength of 405nm, with C...

Embodiment 3

[0125] The Cu prepared by embodiment 3. embodiment 1 + Fluorescent probes for Cu in vitro + Two-photon detection of

[0126] (1) Preparation of calibration curve

[0127] Get the Cu prepared in 1mL embodiment 1 + Fluorescent probes, diluted to 2mL with phosphate buffer, each time adding 10μL of equal concentration of Cu + The solution is then excited by two-photon fluorescence with an excitation wavelength of 800 nm, and the fluorescence intensity of the solution is measured. As the added volume increases, the two-photon fluorescence intensity decreases, and is the same as that of single-photon detection, with good linearity, showing good linearity in the range of 5-80 μM, and the lowest detection limit is 4.40 μM ( Figure 7 ).

[0128] The probe of the present invention has two-photon performance, so not only simple single-photon fluorescence detection, but also two-photon fluorescence detection can be performed. In addition, since the fluorescence lifetime changes linea...

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Abstract

The invention discloses a Cu<+> fluorescent probe and application of the Cu<+> fluorescent probe in cell imaging and biosensing and belongs to the technical fields of fluorescent imaging and biosensing. The invention also discloses a preparation method of the Cu<+> fluorescent probe. The preparation method comprises the following steps: firstly synthesizing a Cu<+> ligand with high selectivity on Cu<+>, and then synthesizing a graphite phase carbon nitride quantum dot (GCNQD), wherein the GCNQD and the Cu<+> ligand are combined to construct a novel Cu<+> fluorescent probe. When Cu<+> exists in solution, the Cu<+> fluorescent probe can be combined with Cu<+> to form a fluorescence weakening substance, fluorescence of the probe is quenched, two-photon fluorescence of the probe is reduced, and fluorescence life is also shortened, so that quantitative detection of Cu<+> in cells can be realized.

Description

technical field [0001] The invention relates to a method for detecting Cu + The novel fluorescent probe and its application in cell imaging and biosensing belong to the technical field of fluorescence imaging and biosensing. Background technique [0002] Monovalent copper ion Cu+ is an essential element in life. Due to its excellent redox properties, it can be used as a coenzyme factor and catalyst to participate in energy production, oxygen transport, cell metabolism and signal transduction in the life process. These are all shown to be essential to life processes. [0003] Cu + Dysregulation within cells triggers dysregulation of redox reactions, leading to abnormal production of reactive oxygen species, resulting in oxidative stress. Oxidative stress disrupts normal physiological processes and leads to cell death, it is closely related to aging and many different diseases such as Menkes, Wilson's disease, neurodegenerative diseases like Alzheimer's, Parkinson's and Hu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C09K11/07G01N21/64
CPCC09K11/07G01N21/6402G01N21/6428G01N21/6458G01N2021/6432
Inventor 田阳李伟
Owner EAST CHINA NORMAL UNIV
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