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A technology of Caenorhabditis elegans and analysis method, which is applied in the direction of analytical materials, animal feed, measuring devices, etc., and can solve the problems affecting the application of neurotoxicity of Caenorhabditis elegans, and the inability to obtain concentration-effect curves for data points, etc.
Pending Publication Date: 2021-12-24
TONGJI UNIV
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Problems solved by technology
Traditional methods cannot obtain complete concentration-effect curves due to fewer data points, which affects the application of C. elegans neurotoxicity in environmental toxicology
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Embodiment 1
[0053] Embodiment 1: S1: the cultivation of escherichia coli
[0054] Dissolve 10g of tryptone, 5g of yeast extract, and 10g of sodium chloride in 1L of ultrapure water, adjust the pH value to 7.0 with 1mol / L NaOH, distribute them in Erlenmeyer flasks, sterilize after bandaging, and place in Store at 4°C to obtain LB liquid medium; inoculate the Escherichia coli freeze-dried powder into the above-mentioned Erlenmeyer flask after activation, and incubate at a constant temperature of 37°C for 24 hours. When the color of the liquid medium changes from transparent yellow to earthy yellow, place Store at 4°C until use;
[0055] S2: Caenorhabditis growth medium configuration:
[0056] Dissolve 17g of agar powder, 2.5g of bacteriological peptone, and 3g of NaCl in 1L of ultrapure water, wrap it up and sterilize it, cool to 70°C and add 1mol / L MgSO sterilized respectively 4 , 1mol / L CaCl 2 , 1 mL each of 5 mg / mL cholesterol ethanol solution and 25 mL of K at pH=6.0 2 HPO 4 -KH 2...
Embodiment 2
[0075] Embodiment 2: S1: the cultivation of escherichia coli
[0076] Dissolve 10g of tryptone, 5g of yeast extract, and 10g of sodium chloride in 1L of ultrapure water, adjust the pH value to 7.0 with 1mol / L NaOH, distribute them in Erlenmeyer flasks, sterilize after bandaging, and place in Store at 4°C to obtain LB liquid medium; inoculate the Escherichia coli freeze-dried powder into the above-mentioned Erlenmeyer flask after activation, and incubate at a constant temperature of 37°C for 24 hours. When the color of the liquid medium changes from transparent yellow to earthy yellow, place Store at 4°C until use;
[0077] S2: Caenorhabditis growth medium configuration:
[0078] Dissolve 17g of agar powder, 2.5g of bacteriological peptone, and 3g of NaCl in 1L of ultrapure water, wrap it up and sterilize it, cool to 70°C and add 1mol / L MgSO sterilized respectively 4 , 1mol / L CaCl 2 , 1 mL each of 5 mg / mL cholesterol ethanol solution and 25 mL of K at pH=6.0 2 HPO 4 -KH 2...
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Abstract
The invention discloses a caenorhabditis elegans dyskinesia-based microplate toxicity analysis method, which is established by taking caenorhabditis elegans as a model organism and introducing a 24-pore plate as a video shooting carrier on the basis of a caenorhabditis elegans lethal microplate toxicity analysis method. The method is characterized by comprising the following steps of: performing pollutant toxicity exposure on caenorhabditis elegans by setting a plurality of concentration gradients in a 96-pore plate by adopting a high-throughput method, then transferring the caenorhabditis elegans into the 24-pore plate to perform caenorhabditis elegans movement behavior disorder video shooting, performing statistical analysis on the video by adopting WormLab software to obtain toxicity data, and finally, carrying out nonlinear curve fitting on the data by adopting a least square method so as to obtain a dose-effect curve of a pollutant to the caenorhabditis elegans dyskinesia. The caenorhabditis elegans dyskinesia-based microplate toxicity analysis method disclosed by the invention can be used for rapidly and accurately acquiring the toxicity data of the pollutant to the caenorhabditis elegans dyskinesia, and is an improvement and an expansion on a traditional caenorhabditis elegans exercise behavior toxicity determination method.
Description
technical field [0001] The invention relates to the technical field of toxicity detection of environmental pollutants, in particular to a toxicity analysis method based on a Caenorhabditis elegans motility disorder microplate. Background technique [0002] With the rapid development of industrialization, many compounds have been developed, resulting in more new pollutants in the environment. Among them, the pollution caused by pesticides, heavy metals, antibiotics and other organic compounds is particularly prominent, which is harmful to the environment and human health. pose huge risks. Therefore, the field of research on the detection of the toxicity of environmental pollutants is becoming more and more extensive. [0003] Physical and chemical analysis and biological detection are mostly used in environmental monitoring. Physical and chemical analysis is to quickly analyze and determine the type and concentration of pollutants through chemical and physical means, and pro...
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