Electrochemical imaging of fusion pore openings by electrochemical detector arrays

Ismail Hafez, Kassandra Kisler, Khajak Berberian, Gregor Dernick, Vicente Valero, Ming G. Yong, Harold G. Craighead, Manfred Lindau

Research output: Contribution to journalArticle

111 Scopus citations

Abstract

Opening of individual exocytotic fusion pores in chromaffin cells was imaged electrochemically with high time resolution. Electrochemical detector arrays that consist of four platinum microelectrodes were microfabricated on a glass coverslip. Exocytosis of single vesicles containing catecholamines from a cell positioned on top of the array is detected by the individual electrodes as a time-resolved oxidation current, reflecting the time course of arrival of catecholamine molecules at the electrode surfaces. The signals exhibit low noise and reveal foot signals indicating fusion pore formation and expansion. The position of individual release events is determined from the fraction of catecholamines recorded by the individual electrodes. Simultaneous fluorescence imaging of release of acridine orange from individual vesicles confirmed the electrochemical position assignments. This electrochemical camera provides very high time resolution, spatiotemporal localization of individual fusion pore openings and quantitative data on the flux of transmitter from individual vesicles. Analysis of the amperometric currents employing random walk simulations indicates that the time course of amperometric spikes measured near the cell surface is due to a low apparent diffusion coefficient of catecholamines near the cell surface and not due to slow dissociation from the granular matrix.

Original languageEnglish (US)
Pages (from-to)13879-13884
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume102
Issue number39
DOIs
StatePublished - Sep 27 2005

Keywords

  • Amperometry
  • Exocytosis
  • Transmitter release

ASJC Scopus subject areas

  • Genetics
  • General

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