Far-field super-resolution microscopy is a rapidly emerging method that is opening up opportunities for biological
imaging beyond the optical diffraction limit. We have implemented a Stimulated Emission Depletion (STED)
microscope to image single dye, cell, and tissue samples with 50-80 nm resolution. First, we compare the STED
performance imaging single molecules of several common dyes and report a novel STED dye. Then we apply STED to
image planar cell polarity protein complexes in intact fixed Drosophila tissue for the first time. Finally, we present a
preliminary study of the centrosomal protein Cep164 in mammalian cells. Our images suggest that Cep164 is arranged in
a nine-fold symmetric pattern around the centriole, consistent with findings suggested by cryoelectron tomography. Our
work demonstrates that STED microscopy can be used for superresolution imaging in intact tissue and provides
ultrastructural information in biological samples as an alternative to immuno-electron microscopy.
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