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Manufacturing method and application of immunofluorescence co-localization imaging platform

An immunofluorescence and co-localization technology, applied in fluorescence/phosphorescence, material analysis through optical means, material inspection products, etc., can solve problems such as false positives and experimental results, and achieve good accuracy.

Pending Publication Date: 2020-04-28
SOUTHEAST UNIV
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

Fluorescence immunoassay has good practicability and ease of use, but due to non-specific adsorption, there are often false positives, which have a great impact on the experimental results.

Method used

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  • Manufacturing method and application of immunofluorescence co-localization imaging platform
  • Manufacturing method and application of immunofluorescence co-localization imaging platform
  • Manufacturing method and application of immunofluorescence co-localization imaging platform

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Experimental program
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Embodiment

[0041] The quartz plate was ultrasonically cleaned in piranha solution for 30 min, then rinsed with water and dried. Place the quartz sheet on a spin coater, add an appropriate amount of PMMA dropwise, with a rotation speed of 500rpm, an increase rate of 2, and a rotation time of 9s, then increase the speed to 4000rpm, an increase rate of 5, and a time of 60s; the drying temperature is 180°C , the time is 2 minutes, and then use the evaporation method to plate an 8nm thick chromium layer to make the quartz sheet conductive. Place the quartz sheet coated with photoresist in an electron beam etching machine for patterning treatment, and soak in chromium etching solution, deionized water, developer, fixer and deionized water in sequence to obtain a patterned quartz sheet .

[0042] The chromium of 5nm and the gold of 15nm are sequentially plated on the quartz sheet by evaporation method, and acetone is put in to remove the redundant photoresist and metal layer, and then the quar...

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Abstract

The invention discloses a manufacturing method and an application of an immunofluorescence co-localization imaging platform. A detection platform generates a gold nano array pattern through an electron beam lithography technology, and a surface is coupled with an HER2 (human epidermal growth factor receptor-2) nucleic acid aptamer with a fluorophore FAM through a gold sulfur bond. The platform canspecifically capture exosomes secreted by HER2 high-expression tumor cells; an imaging chain is an HER2 nucleic acid aptamer probe with another fluorophore Cy5; and two probes have fluorescence scintillation characteristics and can be used for super-resolution monomolecular positioning imaging (SMLM). Through an SMLM imaging technology, the captured exosomes can be subjected to super-resolution optical imaging; in combination with a double-color fluorescence co-localization technology, a false positive event can be judged at a single molecular level, and accuracy of a sandwich immunoassay technology is improved.

Description

technical field [0001] The invention relates to the field of immunodetection and fluorescence colocalization, in particular to a preparation method and application of an immunofluorescence colocalization imaging platform. Background technique [0002] Fluorescence immunoassay technology has the characteristics of strong specificity and good sensitivity, and is widely used in biological detection. It is often used to detect biologically active substances with low content, such as proteins and hormones. The principle of sandwich-type immunoassay is to immobilize excess capture antibody on a solid phase carrier, then add the solution containing the corresponding antigen to be detected, wash it after a period of reaction, and then add an appropriate amount of fluorescently labeled antibody to form an "antibody- Antigen-antibody"sandwich structure complex. The protein content in the solution to be tested can be judged according to the fluorescence intensity. Fluorescence immuno...

Claims

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Application Information

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IPC IPC(8): G01N33/53G01N21/64
CPCG01N33/56966G01N21/6458G01N21/6428G01N2021/6432
Inventor 宗慎飞刘云王著元崔一平
Owner SOUTHEAST UNIV
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