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Immunofluorescence chromatography test strip, immunofluorescence chromatography detection system, and method for determining content of substance to be detected in sample

A technology of immunofluorescence and detection test paper, which is applied in the biological field, can solve the problems of high requirements for color filters, reduced analysis sensitivity, and low fluorescence quantum efficiency, and achieve the effects of strong penetration, improved detection sensitivity, and amplified fluorescent signals

Active Publication Date: 2018-01-26
RESEARCH INSTITUTE OF TSINGHUA UNIVERSITY IN SHENZHEN +2
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  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the emission of traditional fluorescent microspheres is in the visible light region (the emission wavelength is less than 780nm). When it is applied in the fields of living organisms, cell or tissue imaging, and in vitro diagnosis, it is easy to cause poor fluorescence penetration and strong background fluorescence.
At the same time, due to the generally low fluorescence quantum efficiency and the relatively close excitation and emission wavelengths (usually around 20nm), the analysis sensitivity is reduced, and the requirements for the color filters required for fluorescence detection are very high.

Method used

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  • Immunofluorescence chromatography test strip, immunofluorescence chromatography detection system, and method for determining content of substance to be detected in sample
  • Immunofluorescence chromatography test strip, immunofluorescence chromatography detection system, and method for determining content of substance to be detected in sample
  • Immunofluorescence chromatography test strip, immunofluorescence chromatography detection system, and method for determining content of substance to be detected in sample

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preparation example Construction

[0089] According to a specific embodiment of the present invention, the preparation method of the polymer fluorescent microspheres in the near-infrared II region may further include: ultrasonically cleaning the obtained near-infrared II region polymer fluorescent microspheres with ethanol and water in sequence.

[0090] According to a specific embodiment of the present invention, a certain amount of polymer fluorescent microspheres in the near-infrared II region obtained by the preparation method of the above-mentioned embodiment of the present invention is dissolved in dichloromethane, and the fluorescence intensity is measured, and calculated according to the standard curve The number of fluorescent dye molecules contained in each particle is about 80,000. Therefore, compared with small-molecule fluorescent dyes, labeling with polymer fluorescent microspheres in the near-infrared II region can greatly amplify the fluorescent signal and improve detection sensitivity.

[0091]...

Embodiment 1

[0104] (1) Synthesis of carboxylated polystyrene microspheres

[0105] Add 190ml of water into a 500ml round-bottomed flask, and stir at 350rpm in a 70°C water bath for half an hour. Then add 16mg SDS as emulsifier, 0.05g sodium bicarbonate as buffer reagent. Stirring was continued for 10 minutes and 8 mL of styrene and 0.8 mL of acrylic acid were added to the flask. After one hour, 0.2 g of potassium persulfate was added, and the polymerization was carried out under nitrogen protection for 18 hours. After the reaction, wash with 2:1 (v / v) ethanol and aqueous solution and centrifuge three times to obtain carboxylated polystyrene microspheres. Figure 5 (A) shown. Carboxylated polystyrene microspheres are dispersed in the SDS aqueous solution that concentration is 0.25% (w / v) then, configure the carboxylated polystyrene microsphere dispersion liquid of 30mg / ml, as Image 6 As shown in (A), and stored in a 4-degree refrigerator.

[0106] (2) Synthesis of carboxypolystyrene ...

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Abstract

The invention provides an immunofluorescence chromatography test strip, an immunofluorescence chromatography detection system, and a method for determining the content of a substance to be detected ina sample. The immunofluorescence chromatography test strip comprises: a body, wherein a sample zone, a binding zone, a detection zone and an adsorption zone which are sequentially connected are defined on the body; a fluorescence microsphere-labeled first antibody, wherein the first antibody is coated in the binding zone, and the first antibody specifically recognizes the substance to be detected; a detection line and a quality control line which are arranged in the detection zone; a second antibody, wherein the second antibody is coated on the detection line, and the second antibody specifically recognizes the substance to be detected; and a third antibody, wherein the third antibody is coated on the quality control line, the third antibody specifically recognizes the first antibody, andthe above fluorescence microsphere is a near-infrared II zone polymer fluorescence microsphere. The immunofluorescence chromatography test strip has the advantages of high accuracy, high sensitivityand high precision.

Description

technical field [0001] The present invention relates to the field of biology. Specifically, the present invention relates to an immunofluorescence chromatography detection test strip, an immunofluorescence chromatography detection system and a method for determining the content of the analyte in a sample. Background technique [0002] As a special functional microsphere, polymer fluorescent microspheres can contain tens of thousands or even hundreds of thousands of fluorescent molecules, which can greatly improve the efficiency of fluorescent labeling, enhance the anti-photobleaching performance of fluorescent molecules, and greatly Improve analytical sensitivity; at the same time, the surface of fluorescent microspheres can be very flexibly modified with various functional groups (such as carboxyl, amino, aldehyde, etc.), which is conducive to covalent coupling with proteins or antibodies and improves the stability and labeling of markers efficiency. At present, fluoresce...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/558G01N33/543G01N33/533
Inventor 王国新廖滔唐梅杰杨晴来赵肃
Owner RESEARCH INSTITUTE OF TSINGHUA UNIVERSITY IN SHENZHEN