Quantum-dot fluorescent encoded microspheres based on fluorescence resonance energy transfer analysis and preparation method thereof

A fluorescence resonance energy and fluorescence encoding technology, applied in the field of multi-component biomarker analysis, can solve the problems of false positives, easy to be photobleached, strong mutual interference, etc., and achieve the effect of ensuring stability and good fluorescence stability.

Pending Publication Date: 2019-10-08
长春国科希莱科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, the small molecule fluorescent encoding technology is very mature, but there are still problems of broad emission peaks, strong mutual interference, and easy to be photobleached. In addition, the detection system is prone to false positives due to the large number of components that interfere with each other. Therefore, the development of fluorescent substances with narrower fluorescence emission peaks and better stability for encoding, and the construction of a unique optical detection system will improve the above problems

Method used

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  • Quantum-dot fluorescent encoded microspheres based on fluorescence resonance energy transfer analysis and preparation method thereof
  • Quantum-dot fluorescent encoded microspheres based on fluorescence resonance energy transfer analysis and preparation method thereof
  • Quantum-dot fluorescent encoded microspheres based on fluorescence resonance energy transfer analysis and preparation method thereof

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specific Embodiment 1

[0033] Specific embodiment one: the preparation of amino-modified quantum dot coded microspheres comprises the following steps:

[0034] S1: Preparation of polystyrene microspheres: Mix 2 mL of styrene monomer, 0.2 mL of divinylbenzene, and 40-60 mg of AIBN, and then add them to the dispersion medium polyethylene 3-5% polyvinyl alcohol solution, at 70 ° C In a constant temperature four-neck flask, react for 8 h under the condition of blowing nitrogen and stirring (stirring speed is 100 r / min). The product was precipitated by centrifugation, washed three times with ethanol and water respectively, and dried in vacuum to obtain polystyrene microspheres. It looks like image 3 It can be seen from the figure that its morphology is spherical, the particle size distribution is narrow, and it is a uniform monodisperse sphere.

[0035]S2: Amination modification of polystyrene microspheres: 1 g of polystyrene microspheres was weighed and dispersed in 10 mL of deionized water to prepar...

specific Embodiment 2

[0038] Specific embodiment two: the preparation of the quantum dot coded microsphere of carboxyl modification comprises the following steps:

[0039] S1: Preparation of polystyrene microspheres: Mix 2 mL of styrene monomer, 0.2 mL of divinylbenzene, and 40-60 mg of AIBN, and then add them to the dispersion medium polyethylene 3-5% polyvinyl alcohol solution. In a four-neck flask with a constant temperature of ℃, react for 8 h under the conditions of blowing nitrogen and stirring (stirring speed is 100 r / min). The product was precipitated by centrifugation, washed three times with ethanol and water respectively, and dried in vacuum to obtain polystyrene microspheres.

[0040] S2: Carboxylation modification of polystyrene microspheres: Weigh 1g of polystyrene microspheres, 25mg of sodium lauryl sulfate, 250mg of potassium persulfate, add 40mL of distilled water and ultrasonically mix evenly; take another 15mL centrifuge tube, Add 4mL methanol, then add 980μL undecylenic acid an...

specific Embodiment 3

[0043] Specific embodiment three: the preparation of the aldehyde-modified quantum dot coded microsphere comprises the following steps:

[0044] S1: Preparation of polystyrene microspheres: Mix 2 mL of styrene monomer, 0.2 mL of divinylbenzene, and 40-60 mg of AIBN, and then add them to the dispersion medium polyethylene 3-5% polyvinyl alcohol solution. In a four-neck flask with a constant temperature of ℃, react for 8 h under the conditions of blowing nitrogen and stirring (stirring speed is 100 r / min). The product was precipitated by centrifugation, washed three times with ethanol and water respectively, and dried in vacuum to obtain polystyrene microspheres.

[0045] S2: Aldehyde modification of polystyrene microspheres: In a 250mL four-neck flask equipped with a stirrer, add 3g polystyrene microspheres and 30mL dichloromethane to fully swell the microspheres for 12h, then add 13mL 1 , 4-dichloromethoxybutane and 1.5mL of tin tetrachloride were reacted at room temperature ...

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Abstract

The invention discloses quantum-dot fluorescent encoded microspheres based on fluorescence resonance energy transfer analysis. The quantum-dot fluorescent encoded microspheres are characterized in that quantum-dot fluorescent encoded polystyrene microspheres are 2-10 mu m in diameter; the surface of the microspheres is modified with functional groups; three or more types of CdSe@ZnS core-shell quantum dots emitting different wavelengths are embedded in the microspheres; the microspheres can emit the wavelength of 500-600 nm and have the particle size of about 2-10 nm. Amino groups are introduced to the surface of the quantum-dot fluorescent encoded microspheres; after chemical and biological modifications are carried out, the microspheres can be used to construct sensitive elements for liquid chips, so that high-throughput high-specificity analytic detection is achieved for biological macromolecules, such as enzymes and proteins.

Description

technical field [0001] The invention belongs to the field of multi-component biomarker analysis, in particular to a quantum dot fluorescence-encoded microsphere based on fluorescence resonance energy transfer analysis and a preparation method thereof. Background technique [0002] With the rapid development of clinical medicine and clinical laboratory science, the demand for fast and efficient multi-index detection technology for multi-component biomarkers is becoming increasingly urgent. Liquid chip technology for multicomponent biomarker detection. Liquid phase chip technology has become one of the most promising high-throughput multi-index detection technologies due to its advantages of large throughput, good flexibility, high sensitivity, and wide dynamic range. At present, the small molecule fluorescent encoding technology is very mature, but there are still problems of broad emission peaks, strong mutual interference, and easy to be photobleached. In addition, the det...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08F212/08C08F212/36C08F8/04C08F8/30C08F8/00C08F8/24C08F257/02C08F220/64G01N33/533G01N33/543
CPCC08F8/04C08F8/30C08F212/08C08F8/00C08F8/24C08F257/02G01N33/54313G01N33/533C08F212/36C08F220/64
Inventor 高玉舟刘珍妮乔善鹏崔继承庄伟
Owner 长春国科希莱科技有限公司
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