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通过安培法检测到的胞吐事件随碳纤维微电极直径的变化情况的不变性。

Invariance of exocytotic events detected by amperometry as a function of the carbon fiber microelectrode diameter.

作者信息

Amatore Christian, Arbault Stéphane, Bouret Yann, Guille Manon, Lemaître Frédéric, Verchier Yann

机构信息

Laboratoire PASTEUR, Département de Chimie, Ecole Normale Supérieure, CNRS UPMC Univ Paris 06, 24 rue Lhomond, 75231 Paris Cedex 05, France.

出版信息

Anal Chem. 2009 Apr 15;81(8):3087-93. doi: 10.1021/ac900059s.

Abstract

Etched carbon fiber microelectrodes of different radii have been used for amperometric measurements of single exocytotic events occurring at adrenal chromaffin cells. Frequency, kinetic, and quantitative information on exocytosis provided by amperometric spikes were analyzed as a function of the surface area of the microelectrodes. Interestingly, the percentage of spikes with foot (as well as their own characteristics), a category revealing the existence of sufficient long-lasting fusion pores, was found to be constant whatever the microelectrode diameter was, whereas the probability of overlapping spikes decreased with the electrode size. This confirmed that the prespike foot could not feature accidental superimposition of separated events occurring at different places. Moreover, the features of amperometric spikes investigated here (charge, intensity and kinetics) were found constant for all microelectrode diameters. This demonstrated that the electrochemical measurement does not introduce significant bias onto the kinetics and thermodynamics of release during individual exocytotic events. All in all, this work evidences that information on exocytosis amperometrically recorded with the usual 7 microm diameter carbon fiber electrodes is biologically relevant, although the frequent overlap between spikes requires a censorship of the data during the analytical treatment.

摘要

不同半径的蚀刻碳纤维微电极已用于对肾上腺嗜铬细胞发生的单个胞吐事件进行安培测量。将安培峰提供的关于胞吐作用的频率、动力学和定量信息作为微电极表面积的函数进行分析。有趣的是,发现有足峰的百分比(以及它们自身的特征),这一类别揭示了存在足够持久的融合孔,无论微电极直径如何都是恒定的,而重叠峰的概率随电极尺寸减小。这证实了峰前足不可能是在不同位置发生的分离事件的偶然叠加。此外,这里研究的安培峰的特征(电荷、强度和动力学)对于所有微电极直径都是恒定的。这表明电化学测量不会对单个胞吐事件期间释放的动力学和热力学引入显著偏差。总而言之,这项工作证明,用通常直径为7微米的碳纤维电极以安培法记录的关于胞吐作用的信息具有生物学相关性,尽管峰之间频繁重叠需要在分析处理期间对数据进行审查。

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