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Journal of Biomolecular Screening
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pH-Insensitive FRET Voltage Dyes

Michael P. Maher

Johnson & Johnson Pharmaceutical Research and Development L.L.C., San Diego, California, mmaher1{at}prdus.jnj.com

Nyan-Tsz Wu

Johnson & Johnson Pharmaceutical Research and Development L.L.C., San Diego, California

Hong Ao

Johnson & Johnson Pharmaceutical Research and Development L.L.C., San Diego, California

Many high-throughput ion channel assays require the use of voltage-sensitive dyes to detect channel activity in the presence of test compounds. Dye systems employing Förster resonance energy transfer (FRET) between 2 membrane-bound dyes are advantageous in combining high sensitivity, relatively fast response, and ratiometric output. The most widely used FRET voltage dye system employs a coumarin fluorescence donor whose excitation spectrum is pH dependent. The authors have validated a new class of voltage-sensitive FRET donors based on a pyrene moiety. These dyes are significantly brighter than CC2-DMPE and are not pH sensitive in the physiological range. With the new dye system, the authors demonstrate a new high-throughput assay for the acid-sensing ion channel (ASIC) family. They also introduce a novel method for absolute calibration of voltage-sensitive dyes, simultaneously determining the resting membrane potential of a cell. (Journal of Biomolecular Screening 2007:656-667)

Key Words: ion channels • high-throughput screening • cell-based assays • Förster resonance energy transfer • membrane potential

This version was published on August 1, 2007

Journal of Biomolecular Screening, Vol. 12, No. 5, 656-667 (2007)
DOI: 10.1177/1087057107302113
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