Imaging Three-Dimensional Single Molecule Orientations

Abstract

Providing previously obscured positional and orientational information, a novel wide-field optical microscope capable of visualizing three-dimensional orientational dynamics of individual room temperature molecules has been developed. Utilizing a single detector, this facile method enables simultaneous observation of all molecular orientations without polarization optics. Such methods not only provide conclusive, nondestructive evidence of single molecule observation, but will also make single molecule orientational studies accessible to a wide range of researchers. Analysis of observed molecular emission patterns not only directly and noninvasively reveals true 3-D orientational dynamics of individual molecules, but also demonstrates that perceived molecular position is strongly dependent on orientation.