stimulated emission depletion microscopy (STED)

A super-resolution technique using a donut-shaped depletion beam and typical Gaussian-shaped excitation beam in an overlapped configuration. The depletion beam is used to prevent fluorophores from fluorescing via the phenomenon of stimulated emission depletion. The non-linear response of fluorophores to increasing depletion beam intensity makes it possible to excite fluorescence from a sub-diffraction limited area at the center of the overlaid beams.

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A Comparison of STED and STORM Super-Resolution Imaging

Stimulated emission depletion microscopy (STED and the related techniques of ground state depletion (GSD and saturated structured illumination (SSIM) are referred to as ensemble focused light imaging techniques, and are based on non-linear optical effects that typically require the application of multiple high-intensity pulsed lasers with specialized modulation filters to control the excitation beam geometry (a technique commonly termed point-spread function engineering). STED instruments utilize a raster-scan imaging scenario similar to a laser-scanning confocal microscope. In contrast, stochastic optical reconstruction microscopy (STORM), as performed using Nikon's N-STORM system, is a single-molecule approach that relies on activation of a limited subset of the overall molecular population to sequentially image and localize individual emitters on a temporal basis. This interactive tutorial explores the similarities and differences of STED and STORM microscopy.