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Method for detecting difference of cell toxicity between atmospheric nano particles and industrial nano particles

A nanoparticle and cytotoxic technology, applied in biochemical equipment and methods, microbiological measurement/testing, fluorescence/phosphorescence, etc., can solve the problems of low accuracy, many influencing factors, carcinogenicity, etc., and achieve less influencing factors , accurate results, and easy to purchase

Inactive Publication Date: 2012-02-08
SHANGHAI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Traditional research methods such as MTT (3-(4,5)-dimethylthiahiazo(-z-yl)-3,5-di-phenytetrazolium romide, thiazolium blue), LDH (lactate dehydrogenase, lactate dehydrogenase) and other evaluation methods exist some Disadvantages, such as the results are not intuitive, there are many influencing factors, the accuracy is not high, the experiment needs to be repeated many times, it takes a lot of time, and the experimental reagents are carcinogenic, etc.

Method used

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  • Method for detecting difference of cell toxicity between atmospheric nano particles and industrial nano particles
  • Method for detecting difference of cell toxicity between atmospheric nano particles and industrial nano particles
  • Method for detecting difference of cell toxicity between atmospheric nano particles and industrial nano particles

Examples

Experimental program
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Effect test

Embodiment 1

[0029] Such as figure 1 As shown, a method for detecting the cytotoxicity difference of atmospheric nanoparticles and industrial nanoparticles, its operation sequence and steps are as follows:

[0030] a. Aspirate the cells digested with trypsin into a centrifuge tube, discard the supernatant, and add 10% fetal bovine serum to make 3 mL of 1×10 5 cells / mL of the cell suspension, and pipette the cells to disperse evenly, inoculate them in a cell culture dish, and place the culture dish in an incubator for 24 hours;

[0031] b. The prepared industrial nano-NiO, ZnO, CeO 2 Particles and atmospheric nanoparticle solutions were mixed with the culture medium to prepare four kinds of 50μg / mL poisoning solutions; use a pipette gun to suck out the waste liquid in the cell culture dish, and wash the cells with 2mL D-hank`s balanced salt solution; Then use a pipette gun to absorb the poisoning solution and drip it into the cell culture dish to poison the cells, and put it into the inc...

Embodiment 2

[0037] In this example, the cell culture and inoculation are the same as in Example 1, and the poisoning process is slightly different. The difference is that a 100 μg / mL poisoning solution is prepared, and the poisoning components are only industrial nano-NiO and nano-CeO 2 particles.

[0038] Such as Figure 7 , Figure 8 Shown are all the pictures taken after observing the cells with an inverted fluorescence microscope described in Example 2, Figure 9 In order to use IPWIN60 to process the pictures to obtain the initial data, the results show that the fluorescence intensity of the industrial nano-NiO particles is stronger, and the industrial nano-CeO 2 The fluorescence intensity of particle poisoning components is weak, but with the increase of poisoning concentration, industrial nano-NiO particles and CeO 2 The fluorescence intensity is stronger than in Example 1.

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Abstract

The invention relates to a method for detecting the difference of cell toxicity between atmospheric nano particles and industrial nano particles. The method comprises the following steps of: a, using a culture medium to prepare a cell suspension, inoculating the cell suspension in a cell culture dish and culturing the culture dish in an incubator for 24 hours; b, mixing a prepared particulate matter solution and the culture medium and preparing into a contaminated solution; using a D-hank's balanced salt solution to clean cells; carrying out contamination on the cells, and culturing the contaminated cells in the incubator for 4 hours; c, adding a mixed solution of a DCFH-DA fluorescent probe and the culture medium, sealing and wrapping with tin foil paper, and culturing the mixed solution in the incubator for 0.5 hour; d, observing the fluorescence intensity and the fluorescence distribution under an inverted fluorescence microscope, and photographing; and e, processing images by fluorescence analysis software to obtain initial data, analyzing and comparing. According to the method, the cell section preparation is fast and simple, and the raw material is easy to buy; by the cell detection method, the result is accurate, the influence factor is fewer, and multiple nano particles can be detected simultaneously; and laboratory reagents are safe and reliable and have no strong toxicity.

Description

technical field [0001] The invention relates to a method for accurately detecting cytotoxicity differences between atmospheric nanoparticles and industrial nanoparticles, and specifically belongs to the technical field of pollution risk assessment of atmospheric particles and industrial nanoparticles. Background technique [0002] Atmospheric particulate matter has become the primary pollutant of urban air in my country, and atmospheric nanoparticles (or atmospheric ultrafine particles) are an important part of atmospheric particulate matter. Industrial nanoparticles refer to particles with at least one dimension of ≦100nm in three-dimensional space. At present, industrial nanoparticles have been widely used in coatings, cosmetics, catalysts, medicine and other fields. During the production and use of industrial nanoparticles, there will be Distributed into the atmosphere intentionally or unintentionally, becoming part of atmospheric nanoparticles. The research on the healt...

Claims

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Application Information

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IPC IPC(8): G01N21/64C12Q1/02
Inventor 吕森林张睿任晶晶易飞郝晓洁
Owner SHANGHAI UNIV
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