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使用荧光电压敏感染料通过流式细胞术检测石房蛤毒素

Flow cytometric detection of saxitoxins using fluorescent voltage-sensitive dyes.

作者信息

Manger Ronald, Woodle Doug, Berger Andrew, Hungerford James

机构信息

Fred Hutchinson Cancer Research Center, Biologics Production Facility, Seattle, WA 98109, USA.

出版信息

Anal Biochem. 2007 Jul 15;366(2):149-55. doi: 10.1016/j.ab.2007.04.010. Epub 2007 Apr 12.

Abstract

By virtue of their ability to block depolarization of nerve cells, the saxitoxins exert the toxic effects associated with paralytic shellfish poisoning and allow for their detection through various methodologies. When veratridine-induced depolarization is followed using voltage-sensitive fluorescent dyes, the presence of these toxic blocking agents can be observed as a decrease in fluorescence of dye-treated nerve cells. Detection using flow cytometry provides for selection of the most responsive population of cultured mouse neuroblastoma (Neuro 2a) cells thereby enhancing assay sensitivity and this approach can be accomplished in real time. The method is demonstrated in preliminary studies using saxitoxin and crude shellfish extracts.

摘要

由于石房蛤毒素能够阻断神经细胞的去极化,它们会产生与麻痹性贝类中毒相关的毒性作用,并可通过各种方法进行检测。当使用电压敏感荧光染料追踪藜芦定诱导的去极化时,这些毒性阻断剂的存在可表现为染料处理的神经细胞荧光减弱。使用流式细胞术进行检测可选择培养的小鼠神经母细胞瘤(Neuro 2a)细胞中反应最灵敏的群体,从而提高检测灵敏度,并且该方法可实时完成。在使用石房蛤毒素和贝类粗提物的初步研究中展示了该方法。

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