Z-polarized confocal microscopy

In light microscopy the transverse nature of the electromagnetic field prec ludes a strongly focused longitudinal field component, thus confining polar ization spectroscopy and imaging to two dimensions (xy). Here we describe a simple confocal microscopy arrangement that optimizes for signal from mole cules with transition dipoles oriented parallel to the optic axis. In the p roposed arrangement, we not only generate a predominant longitudinally (z) polarized focal field, but also engineer the detection scheme in such a way that in a bulk of randomly oriented molecules, the microscope's effective point-spread function is dominated by the contribution of those molecules t hat are oriented along the optic axis. Our arrangement not only implicitly allows for the determination of the orientation of transition dipoles of si ngle molecules in three dimensions, but also highlights the contribution of z-oriented molecules in three-dimensional imaging. (C) 2001 Society of Pho to-Optical Instrumentation Engineers.