Stimulated Emission Depletion Microscopy (STED)

STED imaging is commonly evaluated by directly comparing diffraction-limited confocal images with STED images, where fine structural details become clearly resolved. Under optimized conditions, STED achieves lateral resolutions of 20–50 nm. Multicolor STED enables nanoscale colocalization of distinct molecular species, while time-gated detection can suppress residual peripheral fluorescence and further enhance effective resolution.

STED-FLIM combines super-resolution imaging with fluorescence lifetime contrast. By integrating time-correlated single-photon counting, lifetime information can be acquired at nanoscale spatial resolution. This enables functional imaging beyond structural detail, allowing discrimination of molecular environments or interactions within sub-diffraction regions.

STED-FCS extends STED to fluorescence correlation spectroscopy by reducing the effective observation volume. The smaller detection volume improves spatial confinement of diffusion measurements and enables correlation analysis at higher fluorophore concentrations. This approach provides access to nanoscale molecular dynamics in membranes and other heterogeneous systems.

The MicroTime 200 integrates STED within a fully modular, time-resolved confocal microscope architecture. Rather than limiting super-resolution to a predefined imaging configuration, the platform allows independent control of excitation, depletion, detection, scanning, and timing electronics.

This structural separation enables integration of additional modalities such as FLIM, FCS, AFM correlation, spectral detection, multiphoton excitation, or cryogenic environments without redefining the core system. STED thus becomes part of a broader experimental framework, supporting custom workflows and method development beyond stand-alone super-resolution imaging.