Immunofluorescence stimulated emission depletion microscopy

We report immunofluorescence imaging with a spatial resolution well beyond the diffraction limit. An axial resolution of 50 nm, corresponding to 1/16 of the irradiation wavelength of 793 nm, is achieved by stimulated emission depletion through opposing lenses. We have demonstrated not only that an antibody-tagged label is stable enough to be recorded in this microscopy mode, but also that subdiffraction resolution can be obtained using a standard immunofluorescence preparation.