用电化学细胞计数法对水泡发射器进行定量化学测量,Accounts of Chemical Research

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用电化学细胞计数法对水泡发射器进行定量化学测量
Accounts of Chemical Research ( IF 16.4 ) Pub Date : 2016-09-13 00:00:00 , DOI: 10.1021/acs.accounts.6b00331
Xianchan Li ₁ , Johan Dunevall ₂ , Andrew G. Ewing _{1,

2}

Affiliation

电化学细胞计数法为我们研究存储在纳米囊泡中的递质分子的化学和化学存储能力增加了新的维度。该方法涉及在电极表面上囊泡的吸附和随后的破裂，在此期间电活性内容物被定量地氧化（或还原）。测得的电流使我们能够使用法拉第定律对囊泡中的分子数量进行计数，并将其与在通信细胞中发生单胞吐事件时释放的分子数量相关联。此方法的原始格式包括毛细管电泳分离步骤，以单独寻址每个囊泡，但是我们最近发现，细胞囊泡倾向于吸附到碳电极上，并自发地以及随机破裂，主要发生单个囊泡事件。尽管这种影响可能不是该过程的重要组成部分，但仍研究了这种称为碰撞电化学细胞计数法的方法，囊泡破裂似乎是在囊泡与电极之间的界面处，并且可能是由电穿孔驱动的。孔径和内含物电解速率是孔径和囊泡中蛋白核的存在的函数。在不含蛋白质的模型脂质体中，随着柔软的纳米颗粒撞击电极并且其内含物被氧化，事件会迅速出现。看来装饰小泡表面的蛋白质对于保持与电极的间隙很重要，并且当该间隙闭合时，发生电穿孔。事件响应时间的模型表明所形成的孔足够小，因此我们可以在纳米尖端电极上进行这些测量，并且已使用这种方法以一种称为细胞内冲击电化学细胞计数法的方式来量化活细胞中的囊泡含量。

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Quantitative Chemical Measurements of Vesicular Transmitters with Electrochemical Cytometry

Electrochemical cytometry adds a new dimension to our ability to study the chemistry and chemical storage of transmitter molecules stored in nanometer vesicles. The approach involves the adsorption and subsequent rupture of vesicles on an electrode surface during which the electroactive contents are quantitatively oxidized (or reduced). The measured current allows us to count the number of molecules in the vesicles using Faraday’s law and to correlate this to the amount of molecules released when single exocytosis events take place at communicating cells. The original format for this method involved a capillary electrophoresis separation step to singly address each vesicle, but we have more recently discovered that cellular vesicles tend to adsorb to carbon electrodes and spontaneously as well as stochastically rupture to give mostly single vesicle events. This approach, called impact electrochemical cytometry, even though the impact is perhaps not the important part of this process, has been studied and the vesicle rupture appears to be at the interface between the vesicle and the electrode and is probably driven by electroporation. The pore size and rate of content electrolysis are a function of the pore diameter and the presence of a protein core in the vesicles. In model liposomes with no protein, events appear extremely rapidly as the soft nanoparticles impact the electrode and the contents are oxidized. It appears that the proteins decorating the surface of the vesicle are important in maintaining a gap from the electrode and when this gap is closed electroporation takes place. Models of the event response times suggest the pores formed are small enough so we can carry out these measurements at nanotip electrodes and we have used this to quantify the vesicle content in living cells in a mode we call intracellular impact electrochemical cytometry.

更新日期：2016-09-13

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