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Recent advances in superresolution techniques have enabled scientific discoveries in multiple disciplines. Stimulated emission depletion (STED) fluorescent microscopy invented by Stefan Hell is one of the most promising superresolution techniques, breaking the diffraction limit in video-rate live-cell imaging. STED achieves superresolution by transiently switching off fluorophores located at the outer rim of a focal spot by de-excitation through stimulated emission. Unlike electron microscopes, STED does not require a vacuum in which to operate. This enables superresolution live cell imaging in an unperturbed environment.
Resolution comparison of confocal and STED microscopes for filopodial actin filaments of gastric cancer cells.
We have built a continuous-wave (CW) STED system that reaches 50 nm resolution for biological samples. We have used this system to image live neurons, cancer cells, and stem cells. Our goal is to use this STED system to faciliate the studies of molecular localization and dynamics in living cells.
Comparison of confocal and STED images of 20 nm yellow-green fluorescent particles.