Circulating tumor cell detection method based on SERS (Surface-enhanced Raman Scattering)

A technology of tumor cells and detection methods, which is applied in the direction of measuring devices, instruments, and material analysis through optical means, and can solve problems such as uncontrollable cell identification and detection, poor reproducibility and reliability of SERS detection results, and loss of cell information.

Inactive Publication Date: 2018-11-23
GUANGDONG MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the prior art, people have used nano-SERS probes to treat living cells for SERS detection, such as introducing active nano-materials into cancer cells, and using surface-enhanced Raman spectroscopy to identify cancer cells provides an effective detection method (patent Publication No. CN103487425A), but the preparation and introduction of active nanoparticles is a cumbersome and high-cost process. The introduction of nanomaterials by electroporation may cause damage to living cells, resulting in the loss of cell information, and the identification and detection of cells. Uncontrollable factors, and due to the complexity of the biological cell system itself, and the interaction between nanomaterials and cell biological systems are always in dynamic changes, the reproducibility and reliability of SERS detection results are relatively poor, resulting in SERS technology Has not been widely used as a tool by biological scholars as fluorescence technology

Method used

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  • Circulating tumor cell detection method based on SERS (Surface-enhanced Raman Scattering)
  • Circulating tumor cell detection method based on SERS (Surface-enhanced Raman Scattering)
  • Circulating tumor cell detection method based on SERS (Surface-enhanced Raman Scattering)

Examples

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Embodiment example 1

[0061] Differentiate tumor cells.

[0062] Take the purchased liver cancer cell line (SMMC7721), laryngeal cancer cell line (Hep2), cervical cancer cell line (Hela), human neuroblastoma cell line (SH-SY5Y), breast tumor cell line (MCF-7), lung cancer cell line (NCI- H446) were respectively inoculated in cell culture plates, after culture, digested with trypsin in the culture medium, then put into a centrifuge tube and centrifuged at 1500 rpm for 5 minutes, poured out the upper layer of trypsin, then added PBS to wash, centrifuged again, and repeated 3 times, the cell suspension was made, and then 2 μl of the solution was added dropwise on the metal silver nanofilm substrate (purchased from Yunyang Photoelectric Co., Ltd., China), and the surface-enhanced Raman spectrum was detected by a confocal Raman spectrometer with a detection range of 600 ~1800cm -1 , the excitation wavelength is 785nm, the sampling time is 2s, and the excitation light power is 1mw. use

[0063] The Va...

Embodiment example 2

[0065] Determination of white blood cells

[0066] Take an appropriate amount of human blood sample, add an appropriate amount of EDTA anticoagulant and hemolysin into the centrifuge tube containing the blood sample, let it stand at room temperature for 15 minutes, and after the red blood cells are dissolved, use 1500 rpm to centrifuge for 5 minutes and directly pour the upper layer, add After mixing with PBS shaker, centrifuge, pour off the supernatant liquid, and then add PBS to resuspend to prepare white blood cell suspension. Then 2 μl of the solution was added dropwise on the metal silver nano film substrate (purchased from Yunyang Photoelectric Co., Ltd., China), and the surface-enhanced Raman spectrum was detected by a confocal Raman spectrometer, with a detection range of 600-1800cm-1 and an excitation wavelength of 785nm. The sampling time is 2s, and the excitation light power is 1mw.

[0067] Use VancouverRamanAlgorithm software to perform 5th-order polynomial proce...

Embodiment example 3

[0069] Determination of red blood cells

[0070] Sample red blood cell separation method: After taking an appropriate amount of human blood samples, use a 15ml centrifuge tube, add 3ml of separation liquid, carefully draw the blood sample with a straw and add it to the separation liquid, and let it stand for 30-40 minutes. After centrifugation, the centrifuge tube is divided into four layers from top to bottom at this time, the lowermost red blood cell layer is removed, and PBS is added to resuspend to prepare white blood cell suspension. Then 2 μl of the solution was added dropwise on the metal silver nano film substrate (purchased from Yunyang Photoelectric Co., Ltd., China), and the surface-enhanced Raman spectrum was detected by a confocal Raman spectrometer, with a detection range of 600-1800cm-1 and an excitation wavelength of 785nm. The sampling time is 2s, and the excitation light power is 1mw.

[0071] Use VancouverRamanAlgorithm software to perform 5th-order polynom...

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Abstract

The invention belongs to the field of medical detection, and particularly relates to a circulating tumor cell detection method based on SERS (Surface-enhanced Raman Scattering). The circulating tumorcell detection method based on SERS disclosed by the invention comprises the following steps of a, putting a solution containing circulating tumor cells on a metal nano-film substrate; b, detecting the SERS by a confocal Raman spectrometer; c, building a cell SERS discrimination model; d, distinguishing the collected Raman spectrum cell variety by the built discrimination model. According to the method, the SERS with strong signals and succinct spectral peak is used for tumor cell authentication. The method can be used for circulating tumor cell authentication, cancer early screening, therapeutic effect evaluation on cancer patients, anti-tumor medicine development and the like, and belongs to a novel technological means for clinical cancer auxiliary diagnosis. The method has the advantages that the tumor cells are not marked or damaged; the sensitivity is high; the repeated detection can be realized, and the like.

Description

technical field [0001] The invention belongs to the field of medical detection, and in particular relates to a SERS-based detection method for circulating tumor cells. Background technique [0002] According to the statistics of the World Health Organization, more than 12 million people worldwide are diagnosed with cancer every year, and about 7.6 million people die of cancer. No matter in developed or developing countries, cancer is one of the main killers that cause people's death. Early screening can significantly improve the survival rate of cancer patients, however, tumors in the body cavity usually require histopathological examination, that is, a needle biopsy is required. As a destructive operation, this method is difficult to achieve early diagnosis of cancer, and may cause complications such as tissue bleeding and infection in the patient's body. Therefore, it is of great significance to develop new methods for cancer screening and diagnosis that are nondestructiv...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/65
CPCG01N21/658
Inventor 李绍新张延娇方向林祁敏倪
Owner GUANGDONG MEDICAL UNIV
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