Takahashi, Y., Shevchuk, A. I., Novak, P., Babakinejad, B., Macpherson, Julie V., Unwin, Patrick R., Shiku, H., Gorelik, J., Klenerman, D., Korchev, Y. E. and Matsue, T. (2012) Topographical and electrochemical nanoscale imaging of living cells using voltage-switching mode scanning electrochemical microscopy. Proceedings of the National Academy of Sciences of the United States of America, Volume 109 (Number 29). pp. 11540-11545. doi:10.1073/pnas.1203570109

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Abstract

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.

Item Type:	Journal Article
Divisions:	Faculty of Science > Chemistry
Journal or Publication Title:	Proceedings of the National Academy of Sciences of the United States of America
Publisher:	National Academy of Sciences
ISSN:	0027-8424
Official Date:	17 July 2012
Dates:	Date Event 17 July 2012 Published
Volume:	Volume 109
Number:	Number 29
Page Range:	pp. 11540-11545
DOI:	10.1073/pnas.1203570109
Status:	Peer Reviewed
Publication Status:	Published
Access rights to Published version:	Restricted or Subscription Access
Funder:	Japan Society for the Promotion of Science (JSPS), Engineering and Physical Sciences Research Council (EPSRC), Chemical and Biological Program of the National Measurement System of the UK Department of Business, Innovation, and Skill, European Research Council (ERC)
Grant number:	22245011 (JSPC); ERC–2009– AdG2471143–QUANTIF (ERC)

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