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Cell probe complex for detecting human breast cancer cells MCF (Michigan cancer foundation)-7 based on Raman spectrum and method for preparing cell probe complex

A technology for detecting human breast cancer cells and Raman spectroscopy, applied in the field of cell detection, can solve problems such as cell destruction and invasiveness, and achieve the effects of strong specificity, high sensitivity, and high Raman signal enhancement ability

Inactive Publication Date: 2012-08-01
NANJING NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Current technologies for single-cell research mainly include fluorescence spectroscopy, scanning probe microscopy, microfluidic technology, and capillary electrophoresis, but most of them are invasive and can damage cells.

Method used

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  • Cell probe complex for detecting human breast cancer cells MCF (Michigan cancer foundation)-7 based on Raman spectrum and method for preparing cell probe complex
  • Cell probe complex for detecting human breast cancer cells MCF (Michigan cancer foundation)-7 based on Raman spectrum and method for preparing cell probe complex
  • Cell probe complex for detecting human breast cancer cells MCF (Michigan cancer foundation)-7 based on Raman spectrum and method for preparing cell probe complex

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Dissolve the aptamer (S2.2, sequence 5'-GCA GTT GAT CCT TTG GAT ACC CTG G-3') in 0.1 mol / L phosphate buffer (PBS, pH 7.4) at a concentration of 300 ng / mL ; the NaAuCl of 1mol / L 4 and 1mol / L AgNO 3 After the solution is mixed evenly at a volume ratio of 1:1, it is added to the nucleic acid aptamer solution, and the Au 3+ 、Ag + The mass ratio of base and base is 1:1:5. Stir the mixture continuously at 4°C for 0.5–2 hours, store it in the dark for 12 hours, and transfer it to a 1cm×1cm quartz cuvette In this method, the Au-Ag-aptamer nanocomposite was obtained by irradiating for 30 minutes under a UV lamp with a wave number of 254 nm; a 0.1 mol / L Rh 6G methanol solution was prepared and added to the Au-Ag-aptamer nanocomposite dispersion In the solution, control the final concentration of Rh 6G to 1 μmol / L, stir at 4 °C for 1 h in the dark, centrifuge at 10,000 rpm for 10 min, and wash the precipitate with 0.1 mol / L PBS solution (pH 7.4) for a total of 3 times , to rem...

Embodiment 2

[0036] Dissolve the aptamer (S2.2, sequence 5'-GCA GTT GAT CCT TTG GAT ACC CTG G-3') in 0.1 mol / L phosphate buffer (PBS, pH 7.4) at a concentration of 300 ng / mL ; the NaAuCl of 1mol / L 4 and 1mol / L AgNO 3 After the solution is mixed uniformly at a volume ratio of 2:1, it is added to the nucleic acid aptamer solution, and the Au 3+ 、Ag + The mass ratio of base and base is 2:1:5, and the mixture is continuously stirred at 4°C for 0.5–2 hours, and stored in a dark place away from light for 12 hours, and then transferred to a 1cm×1cm quartz cuvette In this method, the Au-Ag-aptamer nanocomposite was obtained by irradiating for 30 minutes under a UV lamp with a wave number of 254 nm; a 0.1 mol / L Rh 6G methanol solution was prepared and added to the Au-Ag-aptamer nanocomposite dispersion In the solution, control the final concentration of Rh 6G to 1 μmol / L, stir at 4 °C for 1 h in the dark, centrifuge at 10,000 rpm for 10 min, and wash the precipitate with 0.1 mol / L PBS solution (...

Embodiment 3

[0038] Dissolve the aptamer (S2.2, sequence 5'-GCA GTT GAT CCT TTG GAT ACC CTG G-3') in 0.1 mol / L phosphate buffer (PBS, pH 7.4) at a concentration of 300 ng / mL ; the NaAuCl of 1mol / L 4 and 1mol / L AgNO 3 After the solution is mixed uniformly at a volume ratio of 4:1, it is added to the nucleic acid aptamer solution, and the Au 3+ 、Ag + The mass ratio of base and base is 4:1:5, and the mixture is continuously stirred at 4°C for 0.5–2 hours, and stored in the dark for 12 hours, then transferred to a 1cm×1cm quartz cuvette In this method, the Au-Ag-aptamer nanocomposite was obtained by irradiating for 30 minutes under a UV lamp with a wave number of 254 nm; a 0.1 mol / L Rh 6G methanol solution was prepared and added to the Au-Ag-aptamer nanocomposite dispersion In the solution, control the final concentration of Rh 6G to 1 μmol / L, stir at 4 °C for 1 h in the dark, centrifuge at 10,000 rpm for 10 min, and wash the precipitate with 0.1 mol / L PBS solution (pH 7.4) for a total of 3...

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Abstract

The invention discloses a cell probe complex for detecting human breast cancer cells MCF (Michigan cancer foundation)-7 based on Raman spectrum and a method for preparing the cell probe complex. The cell probe complex is characterized in that nucleic acid aptamer capable of realizing a targeting effect with the MCF-7 cells is used as a template, bimetal alloy nano particles of gold and silver are deposited on a basic group of a DNA (deoxyribose nucleic acid) chain of the nucleic acid aptamer by a photocatalysis method, and Raman signal molecules rhodamine 6G (Rh6G) is fixedly loaded on the surface of each nano particle. The structure of the probe complex is chain-shaped as shown by a photo of a transmission electron microscope, and the probe complex is capable of realizing the targeting effect with the MCF-7 cells. Characteristic absorption peaks of the rhodamine 6G appear on the Raman spectrum under the effect of the MCF-7 cells and the probe complex, and the human breast cancer MCF-7 cells can be effectively, sensitively and specifically detected.

Description

technical field [0001] The invention belongs to the technical field of cell detection, and in particular relates to a cell probe complex for detecting human breast cancer MCF-7 cells by using a surface-enhanced Raman signal and a preparation method thereof. Background technique [0002] Cancer is an important cause of human death. According to the statistics of the World Health Organization, about 7 million people die of various cancer diseases every year. Among women, breast cancer is the cancer with the highest incidence rate, and the incidence rate of breast cancer has shown an obvious upward trend in recent years. In 2008, the incidence rate of breast cancer in the world was as high as 41.4%. Early diagnosis and treatment of breast cancer are of great significance to reduce breast cancer mortality and improve the 5-year survival rate of patients with the disease. [0003] Cancer is a disease caused by the abnormal mechanism of controlling cell growth and proliferation, ...

Claims

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Application Information

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IPC IPC(8): G01N21/65C12N15/115
Inventor 吴萍蔡称心张卉
Owner NANJING NORMAL UNIVERSITY
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