Imaging and orientational properties of single molecule detected by a high NA objective lens

The ability of a high numerical aperture objective lens to detect single molecule emission as a function of molecule’s orientation has been determined numerically and is used to determine the emission image profile of single molecules excited by linearly polarized light. Several mechanism of illumination corresponding to different experimental configurations has been investigated and the effect of an interface in the resulting electric field distribution has been determined. The capacity of a high NA objective lens to collect the fluorescence signal has been determined and has been used to obtain the emission image profile of 1D dipole in full and annular illumination configuration. Result showed the potential use of emission image profile analysis as a tool to obtain 3D orientation of a single molecule.