Abstract
Amperometry is a powerful method to record quantal release events from chromaffin cells and is widely used to assess how specific drugs modify quantal size, kinetics of release, and early fusion pore properties. Surface-modified CMOS-based electrochemical sensor arrays allow simultaneous recordings from multiple cells. A reliable, low-cost technique is presented here for efficient targeting of single cells specifically to the electrode sites. An SU-8 microwell structure is patterned on the chip surface to provide insulation for the circuitry as well as cell trapping at the electrode sites. A shifted electrode design is also incorporated to increase the flexibility of the dimension and shape of the microwells. The sensitivity of the electrodes is validated by a dopamine injection experiment. Microwells with dimensions slightly larger than the cells to be trapped ensure excellent single-cell targeting efficiency, increasing the reliability and efficiency for on-chip single-cell amperometry measurements. The surface-modified device was validated with parallel recordings of live chromaffin cells trapped in the microwells. Rapid amperometric spikes with no diffusional broadening were observed, indicating that the trapped and recorded cells were in very close contact with the electrodes. The live cell recording confirms in a single experiment that spike parameters vary significantly from cell to cell but the large number of cells recorded simultaneously provides the statistical significance.
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Acknowledgements
The authors thank Professor Brian Kim, Professor Gillis, and Dr. Xin Liu for support and advice, Joan S. Lenz for her excellent technical assistance, and Owasco Meat Co., Inc. for providing the bovine adrenal glands. Post-fabrication was performed in Cornell Nanofabrication Facility (CNF), a member of the National Nanotechnology Coordinated Infrastructure (NNCI), which is supported by the National Science Foundation (Grant ECCS-1542081). This work was supported by the National Institutes of Health (NIH) grants R01MH095046 and R43MH109212.
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M.L. is a partner and J.C.R. an employee of the company ExoCytronics, which will develop and commercialize the microchip array platform technology presented here. ExoCytronics has won an SBIR phase I award from the NIH, which is supporting the development of amperometry/FSCV microchip technology.
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This article is part of the special issue on Chromaffin Cells in Pflügers Archiv – European Journal of Physiology
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Huang, M., Delacruz, J.B., Ruelas, J.C. et al. Surface-modified CMOS IC electrochemical sensor array targeting single chromaffin cells for highly parallel amperometry measurements. Pflugers Arch - Eur J Physiol 470, 113–123 (2018). https://doi.org/10.1007/s00424-017-2067-y
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DOI: https://doi.org/10.1007/s00424-017-2067-y