title: Stimulated Emission Depletion Microscopy.
abstract: Despite its short history, diffraction-unlimited fluorescence microscopy techniques have already made a substantial imprint in the biological sciences. In this review, we describe how stimulated emission depletion (STED) imaging originally evolved, how it compares to other optical super-resolution imaging techniques, and what advantages it provides compared to previous golden-standards for biological microscopy, such as diffraction-limited optical microscopy and electron microscopy. We outline the prerequisites for successful STED imaging experiments, emphasizing the equally critical roles of instrumentation, sample preparation, and photophysics, and describe major evolving strategies for how to push the borders of STED imaging even further in life science. Finally, we provide examples of how STED nanoscopy can be applied, within three different fields with particular potential for STED imaging experiments: neuroscience, plasma membrane biophysics, and subcellular clinical diagnostics. In these areas, and in many more, STED imaging can be expected to play an increasingly important role in the future.
authors.affiliation: Royal Institute of Technology (KTH) , Dept Applied Physics, SciLifeLab, 17165 Solna, Sweden.
authors.forename: Hans
authors.initials: H
authors.lastname: Blom
authors.affiliation: Royal Institute of Technology (KTH) , Dept Applied Physics, Albanova Univ Center, 10691 Stockholm, Sweden.
authors.forename: Jerker
authors.identifier: http://orcid.org/0000-0003-3200-0374
authors.initials: J
authors.lastname: Widengren
mesh_terms: D007091:Image Processing, Computer-Assisted; D008856:Microscopy, Fluorescence