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.

Hybrid Structured Illumination Expansion Microscopy Reveals Microbial Cytoskeleton Organization

“…More mature methods, such as stimulated emission depletion microscopy (STED), 5–7 and single-molecule localization microscopy (SMLM, also known as STORM, PALM, etc. ), 8,9 can routinely achieve a spatial resolution better than 50 nm but require substantial technical expertise to execute and often face considerable challenges when imaging multiple channels, densely labeled specimens, or thick specimens more than a few microns from the coverglass substrate.…”

Spaser Nanoparticles for Ultranarrow Bandwidth STED Super‐Resolution Imaging

“…Beyond generation of laser, the physics of stimulated emission has also been utilized in STED microscopy to improve the spatial resolution by inhibiting spontaneous fluorescent emission in specific spatial region through stimulated emission . Unfortunately, current STED microscopy still captures the spontaneous fluorescent emission as the final signal, which suffers the same spectral crosstalk as other methods . In this contribution, we demonstrated a new concept of spaser‐STED through collecting the coherent spaser emission as the imaging signal, rather than the broadband fluorescence.…”

Designing Sub‐2 nm Organosilica Nanohybrids for Far‐Field Super‐Resolution Imaging

“…With the depletion beam, emitters in the periphery could be deexcited via stimulated‐emission depletion and the active emitting region could be confined to the donut center. For practical STED imaging, the typical selection criteria for nanoprobes considered are small size, high fluorescent quantum yield, water solubility, biocompatibility, and photostability, as well as low saturation power required for fluorescence suppression . However, it remains challenging to find a class of STED nanoprobes that fulfill all these requirements in practice.…”