Patent Number: 7,087,416

Title: Detection of transmembrane potentials by optical methods

Abstract: Methods and compositions are provided for detecting changes in membrane potential in membranes biological systems. In one aspect, the method comprises; a) providing a living cell with a first reagent comprising a charged hydrophobic molecule which is typically a fluorescence resonance energy transfer (FRET) acceptor or donor, or is a quencher and is capable of redistributing within the membrane of a biological membrane in response to changes in the potential across the membrane; b) providing the cell with a second reagent that can label the first face or the second face of a biological membrane within the cell; c) detecting light emission from the first reagent or the second reagent. One aspect of this method involves monitoring membrane potential changes in subcellular organelle membranes in a living cells. Another aspect of the invention is the use of certain embodiments of the method for the screening of test chemicals for activity to modulate the activity of a target ion channel. Another aspect of the present invention is a transgenic organism comprising a first reagent that comprises a charged hydrophobic fluorescent molecule, and a second reagent comprising a bioluminescent or naturally fluorescent protein.

Inventors: Tsien; Roger Y. (La Jolla, CA), Gonzalez, III; Jesus E. (San Diego, CA)

Assignee: The Regents of the University of California

International Classification: C12N 13/00 (20060101); C07F 9/06 (20060101); G01N 21/64 (20060101)

Expiration Date: 8/08/02018