Imaging three-dimensional single molecule orientations

Providing previously obscured positional and orientational information, a n ovel wide-field optical microscope capable of visualizing three-dimensional orientational dynamics of individual room temperature molecules has been d eveloped. Utilizing a single detector, this facile method enables simultane ous 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 stu dies accessible to a wide range of researchers. Analysis of observed molecu lar 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.