Fluorescent nanometer composite particle, preparation method and application thereof

A technology of composite particles and fluorescent nanometers, applied in chemical instruments and methods, luminescent materials, etc., can solve the problems of toxic side effects of living cells, unstable fluorescent signals, limited analytical sensitivity, etc., to improve optical stability and prevent fluorescence leakage , to avoid the effect of bleaching

Inactive Publication Date: 2009-11-18
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the traditional fluorescence analysis method has some insurmountable defects
1. Most fluorescent reagents suffer from photobleaching, which leads to unstable fluorescent signals; 2. Fluorescence leakage is serious; 3. Fluorescent reagents and their photolysis products have certain toxic and side effects on living cells; 4. Traditional biomolecular fluorescent labels method, only a few fluorescent molecules can be connected to the active groups of biomolecules, and the analytical sensitivity is limited

Method used

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  • Fluorescent nanometer composite particle, preparation method and application thereof
  • Fluorescent nanometer composite particle, preparation method and application thereof
  • Fluorescent nanometer composite particle, preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] (1) Mix 0.0416g 3-aminopropyltrimethoxysilane and 0.0150g (3.85×10 -5 mol) fluorescein isothiocyanate is dissolved in 0.5315g dehydrated alcohol (the molar ratio of 3-aminopropyltrimethoxysilane, fluorescein isothiocyanate and dehydrated alcohol is 6: 1: 300), obtains mixed liquid;

[0030] (2) Under nitrogen atmosphere, carry out coupling reaction 12h in dark condition (the equation of coupling reaction is as figure 1 shown);

[0031] (3) Add 1.95ml of tetraethyl orthosilicate (the density is 0.933g / ml, the amount of substance is 8.73×10 -3 mol), mass percent concentration is 25% ammoniacal liquor 3.85ml and dehydrated alcohol 35ml (the volume ratio of the three is 1: 2: 18), described tetraethyl orthosilicate and step (1) described isothiocyanate fluorescence The molar ratio of the element is 227:1; place it in a light-tight container, stir and react at 15°C for 24 hours to obtain a suspension;

[0032] (4) Centrifuge the suspension obtained in step (3) (rotating ...

Embodiment 2

[0035] (1) Dissolve 0.0416g 3-aminopropyltrimethoxysilane and 0.0150g fluorescein isothiocyanate in 0.5315g absolute ethanol (3-aminopropyltrimethoxysilane, fluorescein isothiocyanate and The molar ratio of absolute ethanol is 6:1:300), obtains mixed solution;

[0036] (2) Under nitrogen atmosphere, carry out coupling reaction 12h in dark condition;

[0037] (3) Add 2.50ml of tetraethyl orthosilicate (the density is 0.933g / ml, the amount of substance is 11.20×10 -3 mol), mass percentage concentration is 25% ammoniacal liquor 4.20ml and dehydrated alcohol 60ml (three volume ratios are 1: 1.68: 24), described tetraethyl orthosilicate and step (1) described isothiocyanate fluorescence The molar ratio of the element is 290:1; place it in a light-tight container, stir and react at 40°C for 24 hours to obtain a suspension;

[0038] (4) Centrifuge the suspension obtained in step (3) (rotating speed: 8000 rm / min), wash twice with absolute ethanol and distilled water respectively, to...

Embodiment 3

[0041] (1) Dissolve 0.0416g 3-aminopropyltrimethoxysilane and 0.0150g fluorescein isothiocyanate in 0.5315g absolute ethanol (3-aminopropyltrimethoxysilane, fluorescein isothiocyanate and The molar ratio of absolute ethanol is 6:1:300), obtains mixed solution;

[0042] (2) Under an argon atmosphere, carry out the coupling reaction in dark conditions for 12 hours;

[0043] (3) Add 2.30ml of tetraethyl orthosilicate (the density is 0.933g / ml, the amount of substance is 10.30×10 -3mol), mass percent concentration is 25% ammoniacal liquor 3.50ml and dehydrated alcohol 48ml (the volume ratio of the three is 1: 1.52: 20.9), described tetraethyl orthosilicate and step (1) described isothiocyanate fluorescence The molar ratio of the element is 267:1; place it in a closed light-proof container, stir and react at 40°C for 24 hours to obtain a suspension;

[0044] (4) Centrifuge the suspension obtained in step (3) (rotating speed: 8000 rm / min), wash twice with absolute ethanol and dist...

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Abstract

The invention discloses a fluorescent nanometer composite particle, a preparation method and the application thereof. The method includes the following steps: (1) a silane coupling agent and fluorescein isothiocyanate are dissolved in absolute ethyl alcohol to obtain a reaction solution; (2) nitrogen is blown into the reaction solution, and under dark condition the coupled reaction is carried out;and (3) silicon source predecessor and ammonia are added, the ingredients are led into an airtight and lucifugal container and then are stirred to react, and the fluorescent nanometer composite particle can be obtained after centrifugal separation and rinse. The composite particle of the invention has favorable biological safety, can be easily coupled with various biomolecules in a plurality of ways, and can not harm physiological activities; the composite particle can be used for marking biological macromolecules, for detecting trace active substances in a life system, can not harm the physiological activities, and has a considerably important significance on the illustration of the life activity mechanism and the early diagnosis of diseases; and the composite particle has high stabilityand bleaching resistance, and can effectively prevent fluorescence leakage.

Description

technical field [0001] The invention belongs to the field of nanoparticle markers, in particular to a fluorescent nanocomposite particle and its preparation method and application. Background technique [0002] In the field of biological and medical research, exploring and developing highly sensitive non-isotopic detection methods has always been a topic of great concern to researchers, among which fluorescence analysis is an important non-isotopic method. Fluorescently labeled biomolecules (such as proteins) It has played an important role in the fields of biological research and medical research. However, the traditional fluorescence analysis method has some insurmountable defects. 1. Most fluorescent reagents suffer from photobleaching, which leads to unstable fluorescent signals; 2. Fluorescence leakage is serious; 3. Fluorescent reagents and their photolysis products have certain toxic and side effects on living cells; 4. Traditional biomolecular fluorescent labels In...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C09K11/06
Inventor 刘应亮张念椿高燕红
Owner JINAN UNIVERSITY
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