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Surface-enhanced Raman method of modified dimer for detecting and analyzing the life activity state of living cells

A technology for surface-enhanced Raman and life activities, applied in the directions of Raman scattering, material analysis, material excitation analysis, etc. Determining Raman signal measurement benchmarks and other issues

Active Publication Date: 2021-08-17
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are still some defects in the fluorescence detection method: first, many cells have autofluorescence, and the above autofluorescence will interfere with the luminescence of the target fluorescent protein, resulting in ineffective imaging; Weaker, the signal may not be detected effectively; again, when the fluorescent label enters the cell, it may not bind to the target antigen, but free in the cell, resulting in non-specific measurement results, unable to confirm the true location of the target antigen
However, due to the complex structure in the cell, the Raman signal intensity and peak position may be affected by the internal environment of the cell, and the surface-enhanced Raman scattering signal is prone to fluctuations during the measurement process.
None of the existing surface-enhanced Raman scattering measurement methods can determine the measurement benchmark of Raman signals in living cells, which will significantly affect the repeatability and accuracy of Raman signals
In addition, due to the spontaneous aggregation of nanoparticles after entering cells, the surface-enhanced Raman scattering method also has non-specific detection signals.
The existing surface-enhanced Raman scattering method is only applied to the detection of specific protein or organelle Raman spectroscopy, and it is still impossible to confirm whether the substances involved in the life activities in the cell are combined, and it is impossible to realize the positioning imaging and characterization of the specific life activities inside the cell
[0005] To sum up, the existing single-molecule detection methods in living cells are still unable to achieve directional imaging and characterization of substances involved in life activities; surface-enhanced Raman spectroscopy has certain advantages in the field of single-molecule detection in living cells, but its characterization signal is stable. Poor accuracy, lack of baseline information, non-specific signals may appear in the measurement, and has not been applied to the positioning imaging and characterization of specific life activities inside living cells

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  • Surface-enhanced Raman method of modified dimer for detecting and analyzing the life activity state of living cells
  • Surface-enhanced Raman method of modified dimer for detecting and analyzing the life activity state of living cells
  • Surface-enhanced Raman method of modified dimer for detecting and analyzing the life activity state of living cells

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Embodiment 1

[0080] In this example, directional imaging analysis is performed on the life activities of living cells:

[0081] 1. Provide the first type of nanoparticles, the second type of nanoparticles and the third type of nanoparticles

[0082] Silicon dioxide was pre-coated on the surface of gold nanoparticles with a diameter of less than 50 nm to prevent spontaneous aggregation of nanoparticles inside the cells.

[0083] The first type of nanoparticles: DNA template strands are modified on the surface of gold nanoparticles.

[0084] The second type of nanoparticles: RNA polymerase and ribonucleotides are modified on the surface of gold nanoparticles.

[0085] The third type of nanoparticles: human HER2 monoclonal antibody and silicon are modified on the surface of gold nanoparticles, and human HER2 monoclonal antibody can bind to human HER2 protein receptors on organelles.

[0086] 2. Endocytosis of the first type of nanoparticles, the second type of nanoparticles and the third ty...

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Abstract

The present invention proposes a modified dimer surface-enhanced Raman method for detecting and analyzing the life activity state of living cells, including: modifying the first type of nanoparticles with substance A involved in a certain life activity of the cell, and modifying the first type nanoparticles with a substance A involved in the life activity of the cell. The second type of nanoparticles of Substance B is introduced into cells. During the process of life activities, Substance A and B combine, and the nanoparticles modified by the two form dimers, which induce the surface-enhanced pull of the dimer of Substance A and Substance B. Mann scattering, due to the order of magnitude difference between the dimer surface-enhanced Raman scattering intensity and ordinary Raman scattering, the signal intensity is observed by Raman spectrum scanning to confirm the place where life activities occur in living cells, and realize the analysis of living cell life activities. The method of the invention is helpful for accurate detection and analysis of the life activities of living cells, has the advantages of strong detection accuracy, good repeatability and the like, and has scientific research and clinical application value.

Description

technical field [0001] The invention relates to the technical field of living cell imaging detection, and specifically, the invention relates to a modified dimer surface-enhanced Raman method for detecting and analyzing the life activity state of living cells. Background technique [0002] Cells are the basic unit of life activities, and the information changes in cells are the earliest information for the occurrence of diseases. Under the stimulation of different stimuli such as viruses and drugs, studying the location, duration, and changes of reactants of specific life activities inside cells is expected to explain the process of biological disease and healing from the source, and can also quickly verify the effects of drugs. Therefore, the directional imaging research of life activities in living cells is of great significance to the development of biology and pharmacy. [0003] The smallest carrier of life activities in cells is a single molecule. Existing single-mole...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/65
CPCG01N21/658
Inventor 张兴樊傲然李玉璞王海东马维刚常智杰
Owner TSINGHUA UNIV
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