Micro-nano particles detection system and method thereof

A particle detection, micro-nano technology, applied in the direction of nanoparticle analysis, measuring devices, biological particle analysis, etc., can solve the problems of complex samples, heavy operation steps, and lack of adaptability, so as to promote accumulation and increase signal magnification Effect

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

AI Technical Summary

Problems solved by technology

[0009] The above two technical solutions, on the one hand, require special equipment and methods for complicated pretreatment, separation and purification of samples, and heavy operation steps; process is not adaptive

Method used

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  • Micro-nano particles detection system and method thereof
  • Micro-nano particles detection system and method thereof
  • Micro-nano particles detection system and method thereof

Examples

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Effect test

Embodiment 1

[0073] 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, fluorescently labeled aptamers are single-stranded DNA of 20-60 bases, and the diameter of the coil in the sample liquid is less than 5 nanometers, while the diameter of exosomes is 30-150 nanometers; it will specifically recognize CD63 The protein aptamer was 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 culture ...

Embodiment 2

[0078] 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.

[0079] The exosome manipulation method used, as well as the laser, sample chamber, microscope and CCD camera are the same.

[0080] 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

[0084] 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.

[0085] 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 micro-nano particles detection system and a method thereof. The system comprises a heating unit, a sample chamber unit, and a signal acquisition unit, wherein the heating unit is arranged outside the sample chamber unit for heating a sample in the sample chamber unit; micro-nano particle fluid is loaded in the sample chamber unit, the heating unit is used for heating thesample chamber unit, the sample chamber unit generates the thermophoresis effect, the micro-nano particles are gathered at a one side with temperature lower than the micro-nano particle fluid in thesample chamber unit, the signal acquisition unit is used for collecting the correlation information of the micro-nano particles, and corresponding analysis is carried out. The micro-nano detection system performs accumulation on the particles by the thermophoresis effect, gathering and detection can be completed by requiring micro nano particles, pre-treatment and purification of the micro nano particles are not required, the micro-nano particles detection system is generally used for an aptamer and an antibody, and has wide application prospect.

Description

technical field [0001] The invention relates to the technical field of micro-nano particle detection, in particular to a micro-nano particle detection system and method based on thermophoretic effect. 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, clinical medicine 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 instant...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/44G01N1/40G01N21/64
CPCG01N1/4022G01N1/44G01N21/6402G01N21/6428G01N21/6486G01N1/4077G01N15/0612G01N33/54313G01N33/582G01N2015/0038G01N2015/0065G01N2021/6439
Inventor 孙佳姝
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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