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Method for evaluating vascular toxicity of aryl phosphate flame retardant based on high-content imaging system

An aryl phosphoric acid and imaging system technology, applied in image enhancement, image analysis, fluorescence/phosphorescence, etc., can solve problems such as low efficiency, low throughput, and limited information acquisition

Active Publication Date: 2020-04-03
SUN YAT SEN UNIV
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  • Claims
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Problems solved by technology

[0004] Traditional toxicity testing methods are low-throughput, low-efficiency, high-cost, single exposure mode and observation index; and have the disadvantages of limited information acquisition

Method used

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  • Method for evaluating vascular toxicity of aryl phosphate flame retardant based on high-content imaging system
  • Method for evaluating vascular toxicity of aryl phosphate flame retardant based on high-content imaging system
  • Method for evaluating vascular toxicity of aryl phosphate flame retardant based on high-content imaging system

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Embodiment 1

[0083] Aryl phosphate flame retardants--triphenylphosphate (Triphenylphosphate, TPhP) vascular toxicity evaluation (process as follows figure 1 ):

[0084] 1. Acquisition of fluorescent images of blood vessels in zebrafish embryos / larvae treated with triphenyl phosphate

[0085] 1. Chemical treatment and culture of transgenic zebrafish embryos:

[0086] Prepare 21 pre-washed and sterilized 60mm glass dishes, randomly place 25 Tg(kdrl:EGFP) transgenic zebrafish embryos in the glass dishes, set 3 parallel dishes for each dose group, each dish contains 10ml fish water .

[0087] Zebrafish embryos were infected with TPhP at 7 doses of 0, 30, 100, 300, 600, 1200, 2400ug / L within 2 hours of fertilization (2hpf).

[0088] In a constant temperature light incubator at 28°C, light / dark: 14 / 10h cycle culture to 72hpf. Change the water every 24 hours and add 200uL 50XPTU (N-phenylthiourea) to each dish from 24hpf to inhibit the pigment production of zebrafish.

[0089] 2. Prepare aga...

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Abstract

The invention discloses a method for evaluating vascular toxicity of an aryl phosphate flame retardant based on a high-content imaging system. The method comprises the following steps: 1) acquiring azebra fish embryo / juvenile fish vascular fluorescence image treated by the aryl phosphate flame retardant; 2) performing blood vessel image segmentation identification and analysis; and 3) obtaining evaluation and screening results. Vascular images of transgenic zebra fish embryos / juvenile fishes marked by specific vascular fluorescence are collected, and polyvascular parameters are analyzed to realize evaluation and screening of the vascular development toxicity of the aryl phosphate flame retardants. The method comprises the steps that a hardware system is responsible for obtaining blood vessel image data and is composed of a high-content imaging system and a 96-hole glass bottom plate matched with agarose coating; and the software system is used for semi-automatic identification and multi-parameter output analysis of a target blood vessel, and relates to software such as Photoshop, Matlab and GraphPad Prism. The method provided by the invention has the characteristics of high flux and stable result, and can quickly and accurately evaluate and screen the vascular development toxicity of the aryl phosphate flame retardant with multiple parameter indexes.

Description

technical field [0001] The invention relates to an evaluation method of vascular toxicity, belonging to the technical field of toxicity evaluation of toxicological chemicals, in particular to an evaluation method of vascular toxicity of environmental pollutant aryl phosphate flame retardants based on a high-content imaging system. Background technique [0002] With the rapid development of social economy, a large number of chemicals have poured into human production, life and the surrounding environment, and the safety problems that endanger human health have become increasingly prominent. A reasonable comprehensive toxicity test for these chemicals is the basis for risk assessment and risk management. Among them, the evaluation of vascular toxicity of chemicals is currently mainly based on in vitro experiments, and the commonly used animal models for in vivo experiments mainly include mice and chicken embryos. In recent years, imaging methods such as laser confocal have be...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G06T7/00G06T7/11G06T7/62G01N21/64
CPCG01N21/6486G06T7/0012G06T2207/10061G06T2207/10064G06T2207/30101G06T7/11G06T7/62
Inventor 韦艳宏钟霞丽柯炜健王璨
Owner SUN YAT SEN UNIV
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