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Electrically evoking and electrochemically resolving quantal release on a microchip.

Gregory M Dittami1, Richard D Rabbitt

  • 1Department of Bioengineering, University of Utah, Salt Lake City, UT 84118, USA. dittami@eng.utah.edu

Lab on a Chip
|December 22, 2009
PubMed
Summary
This summary is machine-generated.

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This study presents a microchip system for electrically triggering and detecting catecholamine release from cells. The device can resolve single vesicle release and has potential applications in flow cytometry.

Area of Science:

  • Biotechnology
  • Neuroscience
  • Analytical Chemistry

Background:

  • Exocytosis is a fundamental cellular process involving the release of molecules from vesicles.
  • Current methods for studying exocytosis can be limited in temporal resolution and throughput.
  • Rat pheochromocytoma (PC12) cells are a widely used model for studying catecholamine release.

Purpose of the Study:

  • To develop and validate a microchip-based system for electrically evoking and detecting exocytotic catecholamine release.
  • To investigate the influence of cell size and electrical field parameters on exocytosis.
  • To assess the system's capability for high-resolution analysis of exocytosis kinetics.

Main Methods:

  • Utilized a microchip with a microchannel and potentiostat circuit to apply electric fields to PC12 cells.

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  • Electrically stimulated exocytosis by flowing cells through an electric field and applying extracellular voltage pulses.
  • Employed amperometry for simultaneous detection of catecholamine release.
  • Conducted electrical finite element model (FEM) analysis to understand cell-environment electrical interactions.
  • Analyzed single vesicle release, exocytosis kinetics, and spike population statistics.

    • Main Results:

    • Demonstrated exocytosis elicited by both flowing cells through an electric field and applying voltage pulses.
    • FEM analysis showed larger cells experience greater depolarization due to electrical field effects.
    • Observed increased catecholamine release rates from larger cells and cell clusters compared to smaller cells.
    • Successfully resolved single vesicle quantal release (zeptomole range) and characterized vesicle fusion kinetics.
    • Detected catecholamine release from multiple sites, suggesting potential for multi-site exocytosis analysis.

    Conclusions:

    • The developed microchip system effectively evokes and detects electrically stimulated exocytosis in PC12 cells.
    • Cell size and electrical field characteristics significantly influence the depolarization and subsequent exocytotic response.
    • The system offers high sensitivity and resolution for studying exocytosis, capable of analyzing single vesicle events and kinetics.
    • This technology shows promise for integration into flow cytometry for advanced exocytosis analysis and screening.