Michael Joseph Erickson

US Patent:

20040191786, Sep 30, 2004

Filed:

Dec 18, 2003

Appl. No.:

10/467964

Inventors:

David Yue - Baltimore MD, US
Michael Erickson - Baltimore MD, US

International Classification:

C12Q001/68
G01N033/53
G06F019/00
G01N033/48
G01N033/50

US Classification:

435/006000, 435/007100, 702/020000

Abstract:

The invention provides a method for determining a measure of FRET comprising obtaining sequential fluorescent intensity readings from a specimen, such as a cell using three filter cubes. Simple equations manipulate readings from each of the filter sets or cubes to specify a unitless index of FRET called the FRET ratio (FR). FR bears a linear relation to FRET efficiency E. The method also provides for determining the fraction of acceptor-tagged molecules bound by donor-tagged molecules; the relative affinity of binding; and the strength of FRET interactions when all acceptor-tagged molecules are bound by donor. The latter determination enables estimates of the physical distance and/or orientation between interacting fluorophore molecules. The method can be used to detect analytes or inter- or intramolecular interactions. In a preferred aspect, the method is used in an HTS assay to identify modulators of such interactions.