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Related Experiment Videos

pH-Insensitive FRET voltage dyes.

Michael P Maher1, Nyan-Tsz Wu, Hong Ao

  • 1Johnson & Johnson Pharmaceutical Research and Development L.L.C., San Diego, California 92121, USA. mmaher1@prdus.jnj.com

Journal of Biomolecular Screening
|May 23, 2007
PubMed
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Researchers developed new, brighter, pH-insensitive voltage-sensitive dyes for high-throughput ion channel assays. This innovation enables a novel assay for acid-sensing ion channels (ASICs) and improved dye calibration.

Area of Science:

  • Biophysics
  • Molecular Biology
  • Assay Development

Background:

  • High-throughput ion channel assays often rely on voltage-sensitive dyes.
  • Förster resonance energy transfer (FRET) dye systems offer sensitivity, speed, and ratiometric output.
  • Existing FRET voltage dye systems using coumarin donors exhibit pH sensitivity.

Purpose of the Study:

  • To validate a new class of pyrene-based voltage-sensitive FRET donors.
  • To develop a novel high-throughput assay for acid-sensing ion channels (ASICs).
  • To introduce a new method for absolute calibration of voltage-sensitive dyes.

Main Methods:

  • Development and validation of pyrene-based FRET voltage-sensitive dyes.
  • Implementation of a high-throughput assay for ASIC family.

Related Experiment Videos

  • Novel method for simultaneous determination of resting membrane potential and dye calibration.

    • Main Results:

    • The new pyrene-based dyes are brighter and not pH sensitive in the physiological range compared to existing coumarin dyes.
    • A new high-throughput assay for the ASIC family was successfully demonstrated.
    • A novel method for absolute calibration of voltage-sensitive dyes was introduced.

    Conclusions:

    • Pyrene-based FRET dyes offer significant advantages over coumarin-based dyes for voltage-sensitive assays.
    • The developed assay and calibration method advance high-throughput ion channel research.
    • This work provides improved tools for studying ion channel function and drug discovery.