Particle detection system based on microspheres carrier and method thereof

A technology of particle detection and microspheres, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of particle detection that cannot be dissociated, and achieve the effect of convenient operation and promotion of accumulation

Active Publication Date: 2018-09-28
THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0011] The object of the present invention is to provide a particle detection system and method based on a microsphere carrier to overcome the defects in the prior art that cannot detect the accumulation of free particles

Method used

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  • Particle detection system based on microspheres carrier and method thereof
  • Particle detection system based on microspheres carrier and method thereof
  • Particle detection system based on microspheres carrier and method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0078] Incubate exosome samples with fluorescently labeled aptamers, and the selected aptamers are oligonucleotides that can specifically bind to proteins or other small molecular substances screened by the in vitro screening technique SELEX (Systematic Evolution of Ligands by Exponential Enrichment) Fragments, specifically, the fluorescently labeled aptamer is a 40-base single-stranded DNA, and the diameter of the coil in the sample liquid is less than 5 nanometers, while the diameter of the exosome is 30-150 nanometers; it will specifically recognize the CD63 protein Aptamers were applied to exosomes in the culture supernatant of A375 cells (human melanoma cells). Fluorescent groups can be modified at the end of the aptamer by standard means. When the aptamer specifically interacts with the target protein on the surface of the exosome, the exosome is labeled with the fluorescence carried by the aptamer. The exosome sample in this example is the supernatant of the cell cultur...

Embodiment 2

[0083] In this example, using serum samples from patients with cervical cancer, 7 different aptamers were used to detect the abundance of 7 surface proteins (CD63, PTK7, EpCAM, HepG2, HER2, PSA, CA125) of exosomes in serum samples, and Comparison with healthy human serum samples.

[0084] The exosome manipulation method used, as well as the laser, sample chamber, microscope and CCD camera are the same.

[0085] combine Figure 4 As shown, it can be seen that the serum exosomes of this cervical cancer patient highly express CD63 protein, and cancer-related markers PTK7, EpCAM, HepG2, HER2, PSA and CA125, among which CA125 can be used as a traditional cervical cancer marker, and some cervical cancer patients There is high expression of HER2. It is generally believed that tumor markers PTK7 and EpCAM are related to various cancers, HepG2 is mainly specific to liver cancer, and PSA is mainly specific to prostate cancer. However, these tumor markers are not strictly correlated w...

Embodiment 3

[0089] In this embodiment, the micro-nano particles used are non-biological micro-nano particles, specifically fluorescent polystyrene microspheres, the brand is Thermofisher, the diameter is 50 to 200 nanometers, the mass fraction is 0.001%, and it is dissolved in Tween20 containing 0.02%. in aqueous solution. The laser, the sample chamber, the microscope and the CCD camera are all the same as the above-mentioned embodiments 1 and 2.

[0090] combined Figure 5 As shown, all the fluorescent microspheres with different diameters are highly converged at the laser spot, and according to the gray value of the fluorescence measurement and the fluorescence picture, it can be seen that the degree of convergence and the fluorescence intensity increase with the increase of the particle diameter, which is consistent with the work of this embodiment The principle is consistent, that is, large particles tend to aggregate more. This example illustrates that both biological and non-biolo...

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Abstract

The invention relates to a particle detection system based on a microspheres carrier and a method thereof. The system comprises a heating unit and a sample chamber unit, wherein the immunomicrospherefluid combined with target biomolecules is loaded in the sample chamber unit, the immunomicrosphere is prepared by fixing an antibody or an aptamer at the surface of the microsphere, the immunomicrosphere combined with the target biomolecules employs fluorescent label, the heating unit is used for heating the sample chamber unit, the sample chamber unit generates the thermophoresis effect, an exosome is gathered at a low-temperature side in the sample chamber unit, a fluorescence signal labeled at the immunomicrosphere combined with the target biomolecules in the sample room is amplified, andthe detection is carried out. The biomacromolecules such as dissociative protein and nucleic acid are capable of modifying the antibody or the aptamer specifically bind to target protein and nucleic acid on the surface of sphere having a micrometer size, the immunomicrosphere can be obtained for gathering the particles in a free form, and the detection is carried out after gathering.

Description

technical field [0001] The invention relates to the field of micro-nano particle detection, in particular to a particle detection system and method based on a microsphere carrier. Background technique [0002] In the prior art, micro-nano particles are detected to measure particle size, shape, concentration, activity, etc., which are widely used in hematology, immunology, molecular biology and other disciplines. In the prior art, the flow particle detection method is often used to detect micro-nano particles, which is a technology for quantitative analysis and sorting of particles in liquid one by one. The Coulter principle used in the detection refers to: suspension When the particles in the electrolyte pass through the small hole with the electrolyte, they replace the same volume of electrolyte, and in the circuit designed by constant current, the resistance between the two electrodes inside and outside the small hole changes instantaneously, generating a potential pulse, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/533G01N33/543
CPCG01N33/533G01N33/54313
Inventor 孙佳姝刘超田飞
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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