2000-fold parallelized dual-color STED fluorescence nanoscopy.

Bergermann, F.; Alber, L.; Sahl, S. J.; Engelhardt, S.; Hell, S. W.

doi:10.1364/OE.23.000211

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2000-fold parallelized dual-color STED fluorescence nanoscopy.

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Sahl, S. J.
Department of NanoBiophotonics, MPI for biophysical chemistry, Max Planck Society;

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Hell, S. W.
Department of NanoBiophotonics, MPI for biophysical chemistry, Max Planck Society;

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http://www.opticsinfobase.org/oe/viewmedia.cfm?uri=oe-23-1-211&seq=0
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Citation

Bergermann, F., Alber, L., Sahl, S. J., Engelhardt, S., & Hell, S. W. (2015). 2000-fold parallelized dual-color STED fluorescence nanoscopy. Optics Express, 23(1), 211-223. doi:10.1364/OE.23.000211.

Cite as: https://hdl.handle.net/11858/00-001M-0000-0024-6D80-A

Abstract

Stimulated Emission Depletion (STED) nanoscopy enables multi-color fluorescence imaging at the nanometer scale. Its typical single-point scanning implementation can lead to long acquisition times. In order to unleash the full spatiotemporal resolution potential of STED nanoscopy, parallelized scanning is mandatory. Here we present a dual-color STED nanoscope utilizing two orthogonally crossed standing light waves as a fluorescence switch-off pattern, and providing a resolving power down to 30 nm. We demonstrate the imaging capabilities in a biological context for immunostained vimentin fibers in a circular field of view of 20 µm diameter at 2000-fold parallelization (i.e. 2000 “intensity minima”). The technical feasibility of massively parallelizing STED without significant compromises in resolution heralds video-rate STED nanoscopy of large fields of view, pending the availability of suitable high-speed detectors.