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Rapid influenza virus labeling and tracing method based on aggregation-induced emission molecules

A technology for aggregation-induced luminescence and influenza virus, which is applied in the fields of virology and biological sciences, can solve the problems of cumbersome steps, low labeling efficiency, and insufficient brightness, and achieve the effect of simple labeling process, overcoming fluorescence quenching, and little impact on virus activity

Inactive Publication Date: 2019-08-30
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the integration of fluorescent protein genes into the viral genome often affects the genetic stability and infectivity of the virus
Direct labeling with traditional fluorescent molecules such as fluorescein isothiocyanate (FITC) is usually insufficiently bright and prone to photobleaching
Quantum dots must be conjugated to viruses through covalent or non-covalent reactions, which is cumbersome and has low labeling efficiency

Method used

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  • Rapid influenza virus labeling and tracing method based on aggregation-induced emission molecules

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Experimental program
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Effect test

Embodiment 1

[0025] A rapid labeling and tracing method for influenza virus based on aggregation-induced luminescent molecules, the specific steps are as follows:

[0026] 1) Inoculate HeLa cells in good condition into a 15mm glass-bottom culture dish until the cell density reaches 50%;

[0027] 2) Take the molecule n-tetraphenylethylene (TPE) with aggregation-induced emission (AIE) properties10 -6 μM;

[0028] 3) Ultrasound at a power of 50 watts for 20 minutes;

[0029] 4) Centrifuge at 12,000 rpm for 10 minutes, and take 5 microliters of supernatant;

[0030] 5) Take a sterile 0.2 ml centrifuge tube, add 5 microliters of purified influenza virus and 5 microliters of n-tetraphenylethylene supernatant and mix well, so that n-tetraphenylethylene is adsorbed on the surface of influenza virus.

[0031] 6) Take a sterile 1.5 ml centrifuge tube, add 5 microliters of influenza virus labeled with aggregation-induced luminescent molecules and dilute to 1 ml with serum-free medium DMEM;

[003...

Embodiment 2

[0035] A rapid labeling and tracing method for influenza virus based on aggregation-induced luminescent molecules, the specific steps are as follows:

[0036] 1) Inoculate HeLa cells in good condition into a 15mm glass-bottom culture dish until the cell density reaches 50%;

[0037] 2) Take the molecule n-tetraphenylethylene (TPE) with aggregation-induced emission (AIE) properties10 -5 μM;

[0038] 3) Ultrasound with a power of 60 watts for 30 minutes;

[0039] 4) Centrifuge at 13,000 rpm for 10 minutes, and take 10 microliters of supernatant;

[0040] 5) Take a sterile 0.2 ml centrifuge tube, add 10 microliters of purified influenza virus and 10 microliters of n-tetraphenylethylene supernatant and mix well, so that n-tetraphenylethylene is adsorbed on the surface of influenza virus.

[0041] 6) Take a sterile 1.5 ml centrifuge tube, add 10 microliters of influenza virus labeled with aggregation-induced luminescent molecules and dilute to 1 ml with serum-free medium DMEM; ...

Embodiment 3

[0045] A rapid labeling and tracing method for influenza virus based on aggregation-induced luminescent molecules, the specific steps are as follows:

[0046] 1) Inoculate HeLa cells in good condition into a 15mm glass-bottom culture dish until the cell density reaches 50%;

[0047] 2) Take the molecule n-tetraphenylethylene (TPE) with aggregation-induced emission (AIE) properties10 -4 μM;

[0048] 3) Ultrasound at a power of 55 watts for 26 minutes;

[0049] 4) Centrifuge at 12500 rpm for 10 minutes, and take 8 microliters of supernatant;

[0050] 5) Take a sterile 0.2 ml centrifuge tube, add 8 microliters of purified influenza virus and 7 microliters of n-tetraphenylethylene supernatant and mix well, so that n-tetraphenylethylene is adsorbed on the surface of influenza virus.

[0051] 6) Take a sterile 1.5 ml centrifuge tube, add 8 microliters of influenza virus labeled with aggregation-induced luminescent molecules and dilute to 1 ml with serum-free medium DMEM;

[0052...

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Abstract

The invention relates to a rapid influenza virus labeling and tracing method based on aggregation-induced emission molecules. According to the method, the aggregation-induced emission molecules are used as fluorescent labels, and the labeling of influenza viruses by the aggregation-induced emission molecules is realized through electrostatic interaction. Through a laser confocal imaging analysis system, the whole process tracing of an infection process of infecting host cells with the influenza viruses can be realized. The labeling method is simple and efficient, has little influence on virusactivity, and can fully reflect a group dynamic infection behavior of the viruses in a single cell. By combining excellent optical properties of the aggregation-induced emission molecules, a multi-objective, real-time and long-time influenza virus tracing method is established, and a powerful tool is provided for globally and efficiently researching an influenza virus infection mechanism.

Description

technical field [0001] The invention belongs to the field of virology and biological sciences, and relates to a rapid labeling and tracing method for influenza virus based on aggregation-induced luminescent molecules. Background technique [0002] In order to effectively prevent and control the spread of viral diseases, it is particularly important to have a deep understanding of the mechanism of virus infection host and other transmission processes. The interpretation of the dynamic process and mechanism of virus infecting host cells will lay an important theoretical foundation for understanding the pathogenic mechanism of viruses and preventing and treating viral diseases. The process of virus infecting host cells is very complex, often including many stages, multiple pathways and involving different subcellular structures. Single virus tracer technology mainly uses real-time dynamic imaging of fluorescence microscope to monitor the infection path and dynamic behavior of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/533G01N21/64
CPCG01N21/6428G01N33/533G01N33/56983
Inventor 郑斌明东王树超
Owner TIANJIN UNIV
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