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Flow fluorescent micro-sphere standard substance and method for preparing same

A flow fluorescence and standard substance technology, which is applied in the field of preparation of fluorescent microsphere standard substances, can solve the problems of demanding use, storage and transportation, complex preparation process, poor stability, etc., achieve excellent stability and increase specific surface area , the effect of uniform fluorescence intensity

Inactive Publication Date: 2017-03-08
BEIJING HAIAN HONGMENG STANDARD SUSNCE TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because FITC is very sensitive to light, temperature and pH, it is prone to fluorescence quenching and has poor stability, which makes the preparation process of FITC-labeled microspheres complicated, and it is difficult to ensure the monodispersity of particle size and the uniformity of fluorescence intensity. , storage and transportation are demanding

Method used

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  • Flow fluorescent micro-sphere standard substance and method for preparing same
  • Flow fluorescent micro-sphere standard substance and method for preparing same
  • Flow fluorescent micro-sphere standard substance and method for preparing same

Examples

Experimental program
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Effect test

Embodiment 1

[0035] A method for preparing a flow fluorescent microsphere standard substance, comprising the following steps:

[0036] (1) Add 2.2g of polyvinylpyrrolidone, 140mL of ethanol, and 9mL of deionized water into a three-necked flask, add 55g of styrene dissolved in 0.76g of azobisisobutyronitrile and 0.07g of divinylbenzene at one time, stir and dissolve evenly ; Under the protection of nitrogen gas, the temperature was raised to 69° C., and the reaction was carried out for 22 hours; after centrifugal washing and drying, the seed microspheres were obtained, with a particle size of 3.18 μm and a coefficient of variation of 2.14%.

[0037] (2) Weigh 0.4 g of seed microspheres and add them to 100 mL of SDS aqueous solution (0.25% by mass fraction), ultrasonically disperse them to disperse the seed microspheres evenly; dissolve 1.0 mg of ethyl perylene tetracarboxylate in 5 mL of dichloromethane , added to the dispersion system of seed microspheres; after emulsification, shaking, ro...

Embodiment 2

[0040] A method for preparing a flow fluorescent microsphere standard substance, comprising the following steps:

[0041] (1) Add 2.0g of polyvinylpyrrolidone, 120mL of ethanol, and 7mL of deionized water into a three-necked flask, add 50g of styrene dissolved in 0.53g of azobisisobutyronitrile and 0.1g of divinylbenzene at one time, stir and dissolve evenly ; Under the protection of nitrogen gas, the temperature was raised to 70° C., and the reaction was carried out for 20 hours; after centrifugal washing and drying, the seed microspheres were obtained, the particle size was 3.02 μm, and the coefficient of variation was 1.78 / %.

[0042] (2) Weigh 0.5 g of seed microspheres and add them to 50 mL of SDS aqueous solution (0.25% by mass fraction), ultrasonically disperse them to disperse the seed microspheres evenly; dissolve 0.2 mg of diisobutyl perylenedicarboxylate in 5 mL of dichloromethane , added to the dispersion system of seed microspheres; after emulsification, shaking, ro...

Embodiment 3

[0045] A method for preparing a flow fluorescent microsphere standard substance, comprising the following steps:

[0046] (1) Add 1.8g of polyvinylpyrrolidone, 98mL of ethanol, and 5mL of deionized water into a three-necked flask, add 44g of styrene dissolved in 0.41g of azobisisobutyronitrile and 0.05g of divinylbenzene at one time, stir and dissolve evenly ; Under the protection of nitrogen gas, the temperature was raised to 67° C., and the reaction was carried out for 24 hours; after centrifugal washing and drying, the seed microspheres were obtained, the particle size was 3.36 μm, and the coefficient of variation was 3.32%.

[0047] (2) Weigh 1.0 g of seed microspheres and add them to 100 mL of SDS aqueous solution (mass fraction is 0.25%), and ultrasonically disperse them to disperse the seed microspheres evenly; dissolve 5 mg of coumarin 6 in 10 mL of dichloromethane, and add to A dispersion system of seed microspheres; after emulsification, shaking, rotary evaporation, ...

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Abstract

The invention relates to a flow fluorescent micro-sphere standard substance and a method for preparing the same. The method includes steps of (1), preparing seed micro-spheres by the aid of dispersion polymerization processes; (2), dispersing the seed micro-spheres into sodium dodecyl sulfate aqueous solution, dissolving hydrophobic fluorescent dye in swelling agents, adding the hydrophobic fluorescent dye and the swelling agents into dispersion systems of the seed micro-spheres and carrying out emulsification, shaking and rotary evaporation to obtain the flow fluorescent micro-sphere standard substance. The flow fluorescent micro-sphere standard substance and the method have the advantages that the flow fluorescent micro-sphere standard substance is a polystyrene micro-sphere, fluorescent molecules cover the inside of the polystyrene micro-sphere, the particle size of the flow fluorescent micro-sphere standard substance is 3.0-3.5 micrometers, a variation coefficient of the flow fluorescent micro-sphere standard substance is 1%-4%, and the maximum emission wavelength of the flow fluorescent micro-sphere standard substance is 520-540 nm when the flow fluorescent micro-sphere standard substance is excited by 488 nm light; the method is simple, the particle size of the fluorescent micro-sphere standard substance prepared by the aid of the method is strictly monodisperse, and the fluorescent micro-sphere standard substance is uniform in fluorescence intensity and excellent in stability.

Description

technical field [0001] The invention belongs to the technical field of particle preparation and application, and relates to a method for preparing fluorescent microsphere standard substances suitable for flow cytometry. Background technique [0002] Flow cytometry (FCM for short) is a kind of single cell or particle stained by fluorescent pigment under high-speed flow state irradiated with high-energy laser, and measures the intensity of scattered light and emitted fluorescence generated by it, so that the cells (or The modern cell analysis technology for qualitative or quantitative detection of physical, physiological, biochemical, immune, genetic, molecular biology traits and functional status of microparticles can accurately quantitatively analyze and classify individual cells. [0003] Flow cytometry is widely used in the fields of clinical diagnosis and biomedical research. In order to ensure the accuracy of the test results, the flow cytometer needs to use quality cont...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C09K11/02C09K11/06C08F212/08C08F212/36G01N15/14
CPCC09K11/025C08F212/08C09K11/06C09K2211/1011C09K2211/1029G01N15/14C08F212/36
Inventor 李力杜兴科李银窦晓亮李蕴翔
Owner BEIJING HAIAN HONGMENG STANDARD SUSNCE TECH
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