Dielectrophoretic field-flow fractionation system for detection of aquatic toxicants.

Dielectrophoretic field-flow fractionation system for detection of aquatic toxicants.
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DOI:
10.1021/ac801095p
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发表时间:
2008-10-15
影响因子:
7.4
通讯作者:
Gascoyne, Peter
Gascoyne, Peter
中科院分区:
化学1区
文献类型:
--
作者:
Pui-ock, Sittisak;Ruchirawat, Mathuros;Gascoyne, Peter

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介电泳场流分离(dFFF)应用作为一种非接触的方式来感测的变化,在培养的人HL-60细胞的质膜电容和电导率响应于毒物暴露。微图案化的电极在悬浮的细胞上施加电力,当它们移动通过薄室时,细胞以抛物线流动轮廓移动。dFFF峰洗脱时间的相对变化,反映了细胞膜面积和离子渗透性的变化,作为反应指数,在前150分钟的接触8种有毒物质具有不同的单一或混合模式的行动(丙烯腈,放线菌素D,四氯化碳,硫丹,N-亚硝基-N-甲基脲(NMU),百草枯二氯化物,嘌呤霉素,和氧化苯乙烯)。根据特定试剂的作用模式,将dFFF方法与所有毒物的细胞活力测定法以及线粒体电位染料测定法或DNA碱性彗星测定法进行比较。除了低剂量的核酸靶向剂(放线菌素D和NMU),dFFF方法检测所有毒物的灵敏度比其他测定,在某些情况下高达105倍的灵敏度比活力的方法。结果表明,dFFF方法值得进一步研究毒理学的可能适用性。
Dielectrophoretic field-flow fractionation (dFFF) was applied as a contact-free way to sense changes in the plasma membrane capacitances and conductivities of cultured human HL-60 cells in response to toxicant exposure. A micropatterned electrode imposed electric forces on cells in suspension in a parabolic flow profile as they moved through a thin chamber. Relative changes in the dFFF peak elution time, reflecting changes in cell membrane area and ion permeability, were measured as indices of response during the first 150 min of exposure to eight toxicants having different single or mixed modes of action (acrylonitrile, actinomycin D, carbon tetrachloride, endosulfan, N-nitroso-N-methylurea (NMU), paraquat dichloride, puromycin, and styrene oxide). The dFFF method was compared with the cell viability assay for all toxicants and with the mitochondrial potentiometric dye assay or DNA alkaline comet assay according to the mode of action of the specific agents. Except for low doses of nucleic acid-targeting agents (actinomycin D and NMU), the dFFF method detected all toxicants more sensitively than other assays, in some cases up to 105 times more sensitively than the viability approach. The results suggest the dFFF method merits additional study for possible applicability in toxicology.
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