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首页> 美国卫生研究院文献>Proceedings of the National Academy of Sciences of the United States of America >Topographical and electrochemical nanoscale imaging of living cells using voltage-switching mode scanning electrochemical microscopy
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Topographical and electrochemical nanoscale imaging of living cells using voltage-switching mode scanning electrochemical microscopy

机译：使用电压开关模式扫描电化学显微镜对活细胞进行形貌和电化学纳米成像

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We describe voltage-switching mode scanning electrochemical microscopy (VSM-SECM), in which a single SECM tip electrode was used to acquire high-quality topographical and electrochemical images of living cells simultaneously. This was achieved by switching the applied voltage so as to change the faradaic current from a hindered diffusion feedback signal (for distance control and topographical imaging) to the electrochemical flux measurement of interest. This imaging method is robust, and a single nanoscale SECM electrode, which is simple to produce, is used for both topography and activity measurements. In order to minimize the delay at voltage switching, we used pyrolytic carbon nanoelectrodes with 6.5–100 nm radii that rapidly reached a steady-state current, typically in less than 20 ms for the largest electrodes and faster for smaller electrodes. In addition, these carbon nanoelectrodes are suitable for convoluted cell topography imaging because the RG value (ratio of overall probe diameter to active electrode diameter) is typically in the range of 1.5–3.0. We first evaluated the resolution of constant-current mode topography imaging using carbon nanoelectrodes. Next, we performed VSM-SECM measurements to visualize membrane proteins on A431 cells and to detect neurotransmitters from a PC12 cells. We also combined VSM-SECM with surface confocal microscopy to allow simultaneous fluorescence and topographical imaging. VSM-SECM opens up new opportunities in nanoscale chemical mapping at interfaces, and should find wide application in the physical and biological sciences.
机译：我们描述了电压开关模式扫描电化学显微镜（VSM-SECM），其中单个SECM尖端电极用于同时获取活细胞的高质量形貌和电化学图像。这是通过切换施加的电压，从而将法拉第电流从受阻的扩散反馈信号（用于距离控制和地形成像）更改为感兴趣的电化学通量测量来实现的。这种成像方法是可靠的，并且单个纳米级SECM电极易于生产，可用于形貌和活性测量。为了最大程度地减少电压切换时的延迟，我们使用了半径为6.5–100 nm的热解碳纳米电极，该电极可以快速达到稳态电流，对于最大的电极通常不到20毫秒，对于较小的电极通常更快。另外，由于RG值（总探针直径与活性电极直径之比）通常在1.5-3.0范围内，因此这些碳纳米电极也适用于回旋细胞形貌成像。我们首先评估了使用碳纳米电极的恒定电流模式形貌成像的分辨率。接下来，我们进行了VSM-SECM测量，以可视化A431细胞上的膜蛋白并检测PC12细胞的神经递质。我们还将VSM-SECM与表面共聚焦显微镜相结合，以实现同步荧光和地形成像。 VSM-SECM在界面上的纳米级化学制图方面开辟了新的机会，应该在物理和生物科学中得到广泛的应用。

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