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Evaluation method of vascular toxicity of aryl phosphate flame retardants 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 the problems of high cost, single exposure method and observation index, and limited information acquisition

Active Publication Date: 2021-10-19
SUN YAT SEN UNIV
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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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  • Evaluation method of vascular toxicity of aryl phosphate flame retardants based on high-content imaging system
  • Evaluation method of vascular toxicity of aryl phosphate flame retardants based on high-content imaging system
  • Evaluation method of vascular toxicity of aryl phosphate flame retardants 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 the vascular toxicity of aryl phosphate flame retardants based on a high-content imaging system, which comprises the following steps: 1) obtaining vascular fluorescence images of zebrafish embryos / juveniles treated with aryl phosphate flame retardants ; 2) Segmentation, recognition and analysis of blood vessel images; 3) Obtaining evaluation and screening results. By collecting blood vessel images of transgenic zebrafish embryos / juveniles with specific blood vessel fluorescence labeling, and analyzing multiple blood vessel parameters, the evaluation and screening of vascular developmental toxicity of aryl phosphate flame retardants can be realized. The configuration of the method includes: the hardware system is responsible for the acquisition of blood vessel image data, which is composed of a high-content imaging system and a 96-well glass bottom plate coated with agarose; the software system is used for semi-automatic identification of target blood vessels and multi-parameter output analysis, involving Photoshop, Software such as Matlab and GraphPad Prism. The invention has the characteristics of high throughput and stable results, and can quickly and accurately evaluate and screen the vascular developmental toxicity of multi-parameter indexes for aryl phosphate flame retardants.

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 Patents(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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