Fluorescent microspheres and preparation method and fluorescence coding method thereof

A fluorescent microsphere and microsphere technology, applied in chemical instruments and methods, luminescent materials, particle and sedimentation analysis, etc., can solve the problems of fluorescence quenching, inability to meet biochemical detection, affecting the detection of fluorescent signals by instruments, etc. The effect of stability

Pending Publication Date: 2019-11-22
深圳唯公生物科技有限公司
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Problems solved by technology

However, the coding space of fluorescent dyes in these preparation methods is small, and the intensity of labeled fluorescent dyes is low, which cannot meet the needs of biochemical detection.
[0004] In addition, fluorescent dyes are essentially organic substances containing unsaturated bonds, which are easily oxidized by oxygen in air or water, resulting in fluores

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  • Fluorescent microspheres and preparation method and fluorescence coding method thereof
  • Fluorescent microspheres and preparation method and fluorescence coding method thereof
  • Fluorescent microspheres and preparation method and fluorescence coding method thereof

Examples

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preparation example Construction

[0031] A method for preparing fluorescent microspheres according to one embodiment includes the following steps S1-S2.

[0032] Step S1: Activating the microspheres by mixing them with a silane reagent and a solvent to obtain activated microspheres; wherein the silane reagents are epoxy silane reagents, chlorosilane reagents, isocyanate silane reagents, succinic anhydride silane reagents and chlorine At least one of the sulfonylphenylsilane reagents.

[0033] The use of the above-mentioned silane reagents does not require acidic or alkaline conditions to catalyze, and can directly interact with microspheres to activate microspheres under neutral conditions, and form epoxy groups, chlorine groups, isocyanate groups, succinic anhydride groups or Chlorosulfonylphenyl, in addition, it also avoids the problem of poor activation effect caused by ring opening or hydrolysis of these groups under acidic or alkaline conditions; the prepared active microspheres do not need coupling agent...

Embodiment 1

[0077] Surface activation of microspheres: add 30mL deionized water and 1mL of 20mg / mL Fe to a 250mL three-neck flask 3 o 4 The microsphere solution and 1 mL of epoxy silane reagent (3-glycidyloxypropyltriethoxysilane) were stirred at 200 rpm and 50° C. for 24 hours to obtain epoxy-modified activated microspheres. Fe 3 o 4 The particle size of the microspheres is 5 microns.

[0078] Microsphere-coupled fluorescent dye: Add 30 mL of deionized water, the above epoxy-modified activated microsphere and 80 μL of 1 mg / mL PE fluorescent dye into a 250 mL three-neck flask, stir at 200 rpm and 30 °C for 24 h, that is Microspheres with PE fluorescence were obtained.

Embodiment 2

[0080] It is basically the same as Example 1, except that the fluorescent dye is APC, that is, 10 μL, 30 μL, 60 μL, and 120 μL of 1 mg / mL APC dye solution are added to obtain microspheres with four different APC fluorescence intensities.

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Abstract

The invention relates to fluorescent microspheres and a preparation method and a fluorescent coding method thereof. The preparation method comprises the following steps: mixing microspheres with a silane reagent and a solvent for activation to obtain activated microspheres, with the silane reagent being at least one of an epoxy silane reagent, a chlorosilane reagent, an isocyanate silane reagent,a butanedioic anhydride silane reagent and a chlorosulfonyl phenyl silane reagent; and carrying out a coupling reaction on the activated microspheres and fluorescent dye in a solvent to obtain the fluorescent microspheres. According to the preparation method, one or more types of fluorescent dye are coupled to surfaces of the activated microspheres, so that one or more different fluorescence signals can be obtained. Meanwhile, the mass-volume ratio of each fluorescent dye is adjusted, so that one or more microspheres with different fluorescence and different intensities can be obtained, namelyfluorescent coded microspheres are obtained. The prepared fluorescent microspheres have the advantages of large fluorescent dye coding space, high fluorescent dye intensity and the like, and the defect that the fluorescence intensity of fluorescent coded microspheres prepared by traditional preparation methods cannot meet the requirement is avoided.

Description

technical field [0001] The invention relates to the technical field of biomolecular labeling, in particular to a fluorescent microsphere and a method for its preparation and fluorescent coding. Background technique [0002] Suspension microarray technology based on fluorescent microspheres and their fluorescent codes has the ability to simultaneously screen and quantify multiple proteins and cytokines in the same sample, and has high research and application value in the field of disease diagnosis. The core technology of the suspension microarray technology is the fluorescent-encoded microspheres with unique labeling signals during the detection process. In order to make the fluorescent-encoded microspheres applicable to high-throughput multi-indicator detection systems, this requires that the fluorescent-encoded microspheres have high fluorescence intensity, Uniform size, easy surface functionalization, and good biocompatibility. [0003] The traditional preparation of flu...

Claims

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Application Information

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IPC IPC(8): C09K11/02C09K11/06G01N15/14
CPCC09K11/025C09K11/06G01N15/1434
Inventor 褚先锋李为公
Owner 深圳唯公生物科技有限公司
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