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Method for detecting active oxygen content in cells

A technology of active oxygen content and cells, which is applied in the direction of material analysis, measuring devices, and instruments through optical means, can solve problems such as inaccurate detection results, and achieve the effect of overcoming inaccurate measurement and accurate detection results

Active Publication Date: 2017-08-22
NANJING UNIV
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AI Technical Summary

Problems solved by technology

However, in the existing technology, the change of the final fluorescence intensity caused by the change of the cell number is often ignored, resulting in inaccurate detection results

Method used

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  • Method for detecting active oxygen content in cells
  • Method for detecting active oxygen content in cells
  • Method for detecting active oxygen content in cells

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Embodiment 1

[0025] A method for detecting the content of active oxygen in cells, comprising the steps of:

[0026] Step 1: Select the human liver cancer cell line HepG2 as the cell line for the test, observe its growth state under a microscope, remove the original medium, wash the cells with PBS buffer 1 or 2 times, and add 1 mL of trypsin-EDTA to the culture dish Solution, digest the cells; after the cell digestion is complete, add 10mL of fresh medium containing serum to stop the digestion, and blow the obtained cell suspension evenly;

[0027] Step 2, measure the cell density in the cell suspension with a hemocytometer, and seed the cells into a 96-well plate at a density of 10,000 cells per well;

[0028] Step 3: 24 hours after seeding the plate, the cells were exposed to poison, and different concentrations of exposed pollutant arsenic were added to different well plates;

[0029] Step 4: Use the DCFH-DA probe to detect the content of reactive oxygen species (ROS) in cells induced b...

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Abstract

The invention discloses a method for detecting the active oxygen content in cells. The method can be used for obtaining the DCF fluorescence signal value of single live cells corresponding to exposed contaminants under different concentrations. The method for detecting the active oxygen content in cells has the precise detection result; the problem of inaccurate fluorescence intensity measurement due to the cell quantity change caused by contaminant exposure is solved. The detection method provided by the invention has the advantages that the corresponding single cell fluorescence signal value at a certain concentration can be really reflected, so that the injury to the cells due to the contaminant exposure can be really and accurately reflected; the more solid foundation is laid for the application of an in-vitro cell model in the contaminant toxicity evaluation.

Description

technical field [0001] The invention relates to a method for detecting active oxygen content in cells, which belongs to the field of bioactive cell detection. Background technique [0002] In vitro cell model is a technology that simulates the growth environment of organisms in vitro and detects the influence of external stimuli on cells. The use of cells as the test object has the advantages of high sensitivity, greatly shortening the test cycle, and easy to find out the mechanism of action. It not only improves the efficiency of toxicology testing but also reduces the use of animals. It is an important platform in the biological evaluation system. Oxidative damage mechanism is an important mechanism of toxicity for many pollutants. In the process of using in vitro cell models to evaluate the toxicity of pollutants, it is often necessary to detect the content of reactive oxygen species (ROS) in cells to characterize the oxidative damage to the body. The traditional intrac...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
CPCG01N21/6402G01N21/6428G01N21/6452G01N21/6486G01N33/5014G01N33/5067G01N2021/6439
Inventor 吴兵刘苏张徐祥
Owner NANJING UNIV
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