Saturated Forster resonance energy transfer microscopy with a stimulated emission depletion beam: a pathway toward single-molecule resolution in far-field bioimaging

We theoretically demonstrate that the spatial resolution of stimulated emission depletion (STED) microscopy can be substantially enhanced without increasing the intensity of the STED beam. In our scheme, tiny nanobeads codoped with donor and acceptor molecules are used as fluorescent probes, in which Forster resonance energy transfer (FRET) can occur with an similar to 100% efficiency between the donors and acceptors. Enhancement of the depletion of acceptors in the nanobeads with the doughnut-shaped depletion beam can lead to an increase of FRET efficiency in the outer area of the excitation spot, which itself is used for deexciting donor molecules and, consequently, enhancing the optical resolution. (C) 2010 Optical Society of America