Carboxylic fluorescent encoding microsphere and synthetic method thereof

A technology of fluorescent encoding and synthesis method, which is applied in the field of carboxylated fluorescent encoding microspheres and their synthesis, and can solve the problems of reducing

Inactive Publication Date: 2009-03-25
XIAMEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This method of preparing fluorescent microspheres also requires fluorescent molecules to have corresponding reactive groups, and because fluorescent molecules react with functional g

Method used

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  • Carboxylic fluorescent encoding microsphere and synthetic method thereof
  • Carboxylic fluorescent encoding microsphere and synthetic method thereof
  • Carboxylic fluorescent encoding microsphere and synthetic method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1: Synthesis of polystyrene seed microspheres with uniform particle size

[0026]Weigh 0.375g of PVP and 0.25g of AIBN, put the weighed PVP into a 100mL three-neck bottle, add 47.5mL of ethanol to dissolve, add a stirring bar, fix, and stir; add the weighed AIBN to a Add 15mL of St and 0.21mL of MAA to the Erlenmeyer flask, oscillate to dissolve, transfer the solution to a cylindrical dropping funnel, then add it dropwise to a three-necked flask, and install a thermometer; 2 After 10 minutes, the reaction was carried out in an oil bath at 70° C., the stirring speed was 250 rpm, and the reaction was stopped after 24 hours. After cooling, the product was transferred to a 50 mL centrifuge tube and centrifuged at 4000 rpm for 5 min, and then washed three times with ethanol and water successively. The obtained microspheres were dried in a vacuum oven at 70 °C for 24 h. The particle size of the obtained microspheres was 5.7 μm, the dispersion coefficient of the p...

Embodiment 2

[0027] Example 2: Preparation of carboxylated fluorescent microspheres

[0028] Weigh 1 g of 4.73 μm polystyrene seed microspheres, 25 mg of sodium dodecyl sulfate (SDS), potassium persulfate (K 2 S 2 o 8 ) 250mg, fluorescein 5mg, add to 100mL three-neck bottle, add 40mL distilled water, ultrasonic for 5 minutes until the solid is completely dispersed. Take another 15mL centrifuge tube, add 4mL methanol, 980μL undecylenic acid, 10μL divinylbenzene, mix well, pipette 250μL solution into a three-necked bottle, pass N 2 5 to 20 minutes. Stir mechanically (250rpm), place in a 70°C oil bath for 7.5h, stop taking out. The obtained microspheres were centrifuged at 6000 rpm for 4 minutes, washed three times with ethanol, and finally washed three times with deionized water, added with deionized water and stored at 4°C. Compared with the particle size of the seed, the particle size of the obtained microspheres basically does not change, and the uniformity is good, and the fluoresce...

Embodiment 3

[0029] Example 3: Detection of the ability of carboxyl groups on the surface of microspheres to fix biomacromolecules

[0030] Accurately pipette 80 μL of activation buffer solution (10mM sodium acetate, pH5.0) into a 1.5mL centrifuge tube, add 5.0×10 6 Microspheres. After mixing, 0.5 mg and 3.3 mg of freshly prepared NHS and EDC solutions were added, the mixture was incubated at room temperature for 20 min, and centrifuged (10000 rpm × 1 min) to remove unreacted NHS and EDC. Add 250 μL of activation buffer solution to wash once and centrifuge to obtain microspheres; add 500 μL of activation buffer solution, mix well, add 30 μg of hepatitis E antibody (HbeAb), mix well and place it on a horizontal shaker at room temperature for 3.5 hours; then add 500 μL Mix the washing buffer solution evenly, remove the supernatant after centrifugation (10000rpm×1min), and finally add 500 μL preservation buffer solution for preservation.

[0031] Pipette 50 μL of 10 μg / mL phycoerythrin-labe...

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Abstract

The invention discloses a carboxylic fluorescent code microsphere and a synthetic method thereof, which relate to a high molecular material. A simple and easy method for synthesizing the carboxylic fluorescent code microsphere is provided. The carboxylic fluorescent code microsphere is a polystyrene microsphere, the surface of which contains carboxyl and is embedded with organic fluorescent molecules. The particle size of the microsphere is even, ranging from 1 to 10 Mum. By controlling reaction conditions, the microsphere is embedded with fluorescent materials in different types and quantities so as to realize the code of the fluorescent microsphere. A dispersion polymerization method is adopted to synthesize a polystyrene seed microsphere and then the carboxylic microsphere is prepared, and finally the carboxylic fluorescent code microsphere is prepared. The carboxylic fluorescent code microsphere synthesized has even and controllable particle size, stable fluorescent quality and small coefficient of variation and dispersion of fluorescence, thus being applicable to the analysis of liquid phase chips.

Description

technical field [0001] The invention relates to a polymer material, in particular to a carboxylated fluorescent coded microsphere and a synthesis method thereof. Background technique [0002] Liquid phase chip technology is called the chip technology in the post-genome era, also known as flow microbead technology. Since the 1990s, this new technology integrating the fields of molecular biology, immunology, optics and chemistry has been vigorously developed and has become one of the most far-reaching major scientific and technological advances, and it is also the only one approved by the FDA that can be applied to clinical diagnosis. chip technology. The carrier of liquid-phase chip technology is surface-functionalized fluorescent-encoded microspheres. Surface-functionalized fluorescent microspheres are widely used in many fields, especially biomedical fields, due to their strong ability to immobilize biomacromolecules and stable and efficient luminous efficiency. High rese...

Claims

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Application Information

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IPC IPC(8): C09K11/06C08F212/08C08F8/00G01N33/52G01N21/64
Inventor 颜晓梅张志灵潘建波许艳军
Owner XIAMEN UNIV
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