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Fluorescence Imaging of Electrically Stimulated Cells

Johnson & Johnson Pharmaceutical Research and Development, LLC, Raritan, NJ

Janet K. Robertson

Standard MEMS, Inc., Easton, PA

Jeffrey M. Palmer

Richard R. Ryan

Johnson & Johnson Pharmaceutical Research and Development, LLC, Spring House, PA

Adrienne E. Dubin

Johnson & Johnson Pharmaceutical Research and Development, LLC, San Diego, CA

Robert A. Zivin

Johnson & Johnson Pharmaceutical Research and Development, LLC, Raritan, NJrzivin{at}prdus.jnj.com

Designing high-throughput screens for voltage-gated ion channels has been a tremendous challenge for the pharmaceutical industry because channel activity is dependent on the transmembrane voltage gradient, a stimulus unlike ligand binding to G-protein-coupled receptors or ligand-gated ion channels. To achieve an acceptable throughput, assays to screen for voltage-gated ion channel modulators that are employed today rely on pharmacological intervention to activate these channels. These interventions can introduce artifacts. Ideally, a high-throughput screen should not compromise physiological relevance. Hence, a more appropriate method would activate voltage-gated ion channels by altering plasma membrane potential directly, via electrical stimulation, while simultaneously recordingthe operation of the channel in populations of cells. The authors present preliminary results obtained from a device that is designed to supply precise and reproducible electrical stimuli to populations of cells. Changes in voltage-gated ion channel activity were monitored using a digital fluorescent microscope. The prototype electric field stimulation (EFS) device provided real-time analysis of cellular responsiveness to physiological and pharmacological stimuli. Voltage stimuli applied to SK-N-SH neuroblastoma cells cultured on the EFS device evoked membrane potential changes that were dependent on activation of voltage-gated sodium channels. Data obtained using digital fluorescence microscopy suggests suitability of this system for HTS.

Key Words: voltage-gated ion channels • electric field stimulation

Journal of Biomolecular Screening, Vol. 8, No. 6, 660-667 (2003)
DOI: 10.1177/1087057103258546


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