Evaluation method for measuring cell DNA damage caused by soluble heavy metal

A DNA damage and evaluation method technology, applied in the direction of material excitation analysis, fluorescence/phosphorescence, etc., to achieve the effect of reliable data base, fast and accurate simultaneous detection of multiple samples, and less sample volume

Active Publication Date: 2015-09-23
CHINA TOBACCO YUNNAN IND
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  • Summary
  • Abstract
  • Description
  • Claims
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Problems solved by technology

However, the test using γH2AX index to evaluate DNA damage caused by heavy metals is rarely reported at home and abroad.

Method used

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  • Evaluation method for measuring cell DNA damage caused by soluble heavy metal
  • Evaluation method for measuring cell DNA damage caused by soluble heavy metal
  • Evaluation method for measuring cell DNA damage caused by soluble heavy metal

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Experimental program
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Embodiment Construction

[0029] The following are examples of the present invention, but the content of the present invention is not limited thereto.

[0030] 1. Experimental materials: Chinese hamster ovary cells (CHO cells) were provided by Kunming Institute of Zoology, Chinese Academy of Sciences.

[0031] 2. Main experimental equipment: high-content imaging system (ImageXpress MICRO, Molecular Devices); CO 2 Incubator (Thermo Company); secondary biological safety cabinet (Heal Force Company); inverted microscope (TS100-F-HMC type, Nikon Company); 96-well plate, cell culture flask (Corning Company, USA).

[0032] 3. Experimental method:

[0033] (1) Preparation of standard stock solution of hexavalent chromium: weigh 2.9418g K 2 Cr 2 o 7 (Excellent grade pure), after dissolving in water, transfer to a 100ml volumetric flask, dilute with water to the marked line, and shake well. The solubility of this solution is 100mM, ready for use.

[0034](2) CHO cells were planted in a 96-well cell cultur...

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Abstract

The invention relates to an evaluation method for measuring cell DNA damage caused by soluble heavy metal. The method includes the following steps that firstly, tested cells are processed through a soluble heavy metal solution; secondly, the processed tested cells are fixed and are subjected to cell permeabilization; thirdly, the tested cells having been subjected to cell permeabilization are dyed; fourthly, detection is performed through a high-content imaging system and analysis is performed. The evaluation method for measuring the cell DNA damage caused by the soluble heavy metal is convenient to operate, high in sensitivity and high in detection speed and has high specific pertinence, and research on the genetic toxicity of the heavy metal is supplemented and expanded.

Description

technical field [0001] The invention relates to an evaluation method for measuring cellular DNA damage caused by soluble heavy metals by using a high-content imaging system, and belongs to the technical field of environmental ecotoxicology. Background technique [0002] Heavy metals exist in the air, soil, and water that humans depend on for survival, and there are various types, among which mercury, cadmium, lead, chromium, arsenic, etc. have significant biological toxicity, and can enter benthic organisms from water and soil through food chains and other channels , fish, birds and mammals, resulting in biomagnification. At present, heavy metal pollution exists in part of the soil and water in our country. Studies have shown that the toxicity of heavy metals has multi-faceted and multi-level characteristics. However, previous studies have mainly focused on the physical and chemical detection of pollutants. Ecological evaluation of heavy metals. Regarding the toxicity of h...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
Inventor 高茜夭建华黄海涛李雪梅米其利曾婉俐管莹朱洲海
Owner CHINA TOBACCO YUNNAN IND
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