Modified dimer surface enhanced Raman method for detecting and analyzing vital activity state of living cells

Abstract

The invention provides a modified dimer surface enhanced Raman method for detecting and analyzing the vital movement state of living cells. The method comprises the following steps: introducing a first type of nanoparticles modified with a substance A participating in certain vital movement of cells and a second type of nanoparticles modified with a substance B participating in the vital movementof cells into the cells, wherein in the vital movement process, the substance A and the substance B are combined, nanoparticles modified by the substance A and the substance B form a dimer, surface enhanced Raman scattering of the dimer of the substance A and the substance B is induced to be generated, and due to the fact that the dimer surface enhanced Raman scattering intensity is different fromthat of common Raman scattering in order of magnitude, the signal intensity is observed through Raman spectrum scanning, and the occurrence place of the vital movement in the living cells is confirmed so as to realize living cell vital movement analysis. The method is beneficial to accurate detection and analysis of living cell vital movement, has the advantages of high detection accuracy, good repeatability and the like, and has scientific research and clinical application values.

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...

AI Technical Summary

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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