Interferometrical single-molecule localization based on dynamic PSF engineering.

We present a method for interferometric single-molecule localization based on dynamic point spread function (PSF) engineering. By using two galvo mirrors, a hexagonal PSF is constructed and the fluorescent signal under different illumination patterns could be acquired simultaneously. This method was evaluated using simulation, fluorescent nanosphere imaging, and single-molecule imaging. The study indicates a twofold improvement in localization precision while maintaining the same photon budget. This strategy, we believe, is a cost-effective way to increase the resolution of single-molecule localization microscopy.