Rapid determination of the three-dimensional orientation of single molecules

A technique is proposed for determination of the three-dimensional orientation of the transition dipoles of single molecules by use of polarization-sensitive detection of fluorescence through a high-N.A. objective. Molecular orientation can be determined uniquely and rapidly based on the counts from three detectors that are sensitive to different polarizations of light. (C) 2001 Optical Society of America OCIS codes: 180.2520, 260.5430.