Optimizing Dual Fluorescent Analysis to Investigate the Toxicity of AgNPs in E. coli.

Wei Hong, Shaopeng Chen
{"title":"Optimizing Dual Fluorescent Analysis to Investigate the Toxicity of AgNPs in E. coli.","authors":"Wei Hong,&nbsp;Shaopeng Chen","doi":"10.1002/cptx.28","DOIUrl":null,"url":null,"abstract":"<p><p>The ever-increasing use of silver nanoparticles (AgNPs) carries potential ecotoxicological risks. For full risk assessment, E. coli cells harboring a plasmid with a constitutively expressed GFP gene under control of lac promoter (lac::GFP) are extensively utilized. Flow cytometry is an advanced technology usually applied to toxicological research for rapid, efficient, multi-parameter analysis of single cells. However, it is difficult to accurately and sensitively detect the toxicity of nanoparticles with flow cytometry due to the interference of aggregated nanoparticles. In this protocol, dual-fluorescence detection with a propidium iodide (PI)-lac::GFP assay is used to determine the toxicity of AgNPs and successfully discriminate the dead or fragilized bacteria from living bacteria and aggregated nanoparticles. © 2017 by John Wiley & Sons, Inc.</p>","PeriodicalId":72743,"journal":{"name":"Current protocols in toxicology","volume":"73 ","pages":"20.14.1-20.14.13"},"PeriodicalIF":0.0000,"publicationDate":"2017-08-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cptx.28","citationCount":"1","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current protocols in toxicology","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1002/cptx.28","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 1

Abstract

The ever-increasing use of silver nanoparticles (AgNPs) carries potential ecotoxicological risks. For full risk assessment, E. coli cells harboring a plasmid with a constitutively expressed GFP gene under control of lac promoter (lac::GFP) are extensively utilized. Flow cytometry is an advanced technology usually applied to toxicological research for rapid, efficient, multi-parameter analysis of single cells. However, it is difficult to accurately and sensitively detect the toxicity of nanoparticles with flow cytometry due to the interference of aggregated nanoparticles. In this protocol, dual-fluorescence detection with a propidium iodide (PI)-lac::GFP assay is used to determine the toxicity of AgNPs and successfully discriminate the dead or fragilized bacteria from living bacteria and aggregated nanoparticles. © 2017 by John Wiley & Sons, Inc.

优化双荧光分析研究AgNPs在大肠杆菌中的毒性。
银纳米颗粒(AgNPs)的不断增加的使用具有潜在的生态毒理学风险。为了进行全面的风险评估,大肠杆菌细胞被广泛使用,其中含有在lac启动子(lac::GFP)控制下组成表达GFP基因的质粒。流式细胞术是一种用于毒理学研究的先进技术,可对单个细胞进行快速、高效、多参数的分析。然而,由于纳米颗粒聚集的干扰,流式细胞术难以准确、灵敏地检测纳米颗粒的毒性。在该方案中,采用碘化丙啶(PI)-lac::GFP双荧光检测来确定AgNPs的毒性,并成功地将死亡或脆弱的细菌与活细菌和聚集的纳米颗粒区分开来。©2017 by John Wiley & Sons, Inc。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 求助全文
来源期刊
自引率
0.00%
发文量
0
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信