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Early cancer detection method based on single cell level

A cancer detection and single-cell technology, applied in biological testing, measuring devices, material inspection products, etc., can solve the problems of insufficient sensitivity and poor repeatability, and achieve accurate detection results

Pending Publication Date: 2019-01-11
JIANGSU RAYME BIOTECH
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Problems solved by technology

[0003] "Optical Detection of Lactate Release from Single Cells in Tumor Metabolism Analysis" in Analytical Chemistry, Volume 82, 5082-5087, 2010 (Anal.Chem.2010, 82, 5082–5087), an early detection method based on single-cell level Cancer detection method, but the sensitivity of this method is not high enough, and the repeatability is poor, so the present invention further optimizes the test parameters with higher sensitivity and better repeatability

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  • Early cancer detection method based on single cell level
  • Early cancer detection method based on single cell level
  • Early cancer detection method based on single cell level

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Embodiment Construction

[0025] The present invention will be described in detail below in conjunction with the accompanying drawings and specific embodiments.

[0026] A method for early detection of cancer based on a single cell level, comprising the steps of:

[0027] Step 1, the silicon dioxide surface of the nanoprobe tip was silanized in 5% APTES ethanol solution at 60°C for 3 hours, vacuum dried at 120°C for 2 hours, then activated in 5% glutaraldehyde solution at 4°C for 12 hours, and then Incubate in 2mg / ml, 1.8mg / ml, 1.6mg / ml, 1.5mg / ml lactate dehydrogenase solutions for 24 hours at 4°C for immobilization.

[0028] Step 2, select human breast cancer cell line MCF7 as the cell line for the test, observe its growth state under a microscope, remove the original medium, wash the cells 1-2 times with PBS buffer, add 2ml of 0.25% Digest the cells with trypsin solution. After the digestion of the cells is completed, add 2ml of fresh medium containing serum to stop the digestion, suck the suspensio...

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Abstract

The invention discloses an early cancer detection method based on a single cell level. According to the method, lactic dehydrogenase is modified on the tip of a nano-probe, a human breast cancer cellline MCF7 is selected as a cell strain for testing, and the concentration of the extracellular lactic acid of tumor cells can be monitored by catalyzing the conversion of the lactic acid and generating NADH as a fluorescent detection object. The detection method is accurate in detection result of the concentration of the extracellular lactic acid of the cells and is capable of truly reflecting a single cell fluorescence signal value corresponding to a certain concentration, so that the extracellular release of the lactic acid can be reflected truly and accurately. According to the detection method, the metabolism of normal cells and cancer cells can be distinguished from the single cell level, and the method has important significance for understanding a molecular mechanism of cancerationand diagnosis and prognosis of early cancers.

Description

technical field [0001] The invention relates to an early cancer detection method based on a single cell level, belonging to the field of biological cell detection. Background technique [0002] Tumor cells have unique metabolic phenotypes during carcinogenesis. Therefore, the study of tumor metabolism is promising for a large number of applications in oncology, including early cancer diagnosis, disease prognosis, treatment response assessment, and treatment progression. The study of metabolism at the single-cell level is very useful, as it provides opportunities to reveal cellular heterogeneity and decipher complex metabolic interactions, thereby further deciphering various metabolic pathways. Therefore, the biosensing platform can distinguish the metabolism of normal cells and cancer cells from the single-cell level, which is especially important for understanding the molecular mechanism of carcinogenesis and the diagnosis and prognosis of early cancer. [0003] "Optical D...

Claims

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Application Information

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IPC IPC(8): G01N21/64G01N33/50
CPCG01N21/6486G01N33/5008
Inventor 徐艇
Owner JIANGSU RAYME BIOTECH
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