Gated CW-STED microscopy: A versatile tool for biological nanometer scale investigation

[Display omitted] Stimulation emission depletion (STED) microscopy breaks the spatial resolution limit of conventional light microscopy while retaining its major advantages, such as working under physiological conditions. These properties make STED microscopy a perfect tool for investigating dynamic...

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Bibliographic Details
Published in:Methods (San Diego, Calif.) Calif.), 2014-03, Vol.66 (2), p.124-130
Main Authors: Vicidomini, Giuseppe, Coto Hernández, Ivan, d’Amora, Marta, Cella Zanacchi, Francesca, Bianchini, Paolo, Diaspro, Alberto
Format: Article
Language:English
Subjects:
Animals
Calibration
Cercopithecus aethiops
COS Cells
Fluorescent Dyes - chemistry
HeLa Cells
Humans
Limit of Detection
Microscopy, Fluorescence - instrumentation
Microscopy, Fluorescence - methods
Microtubules - ultrastructure
Nanostructures - chemistry
Spatial resolution
Stimulated emission depletion microscopy
Super-resolution microscopy
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Summary:[Display omitted] Stimulation emission depletion (STED) microscopy breaks the spatial resolution limit of conventional light microscopy while retaining its major advantages, such as working under physiological conditions. These properties make STED microscopy a perfect tool for investigating dynamic sub-cellular processes in living organisms. However, up to now, the massive dissemination of STED microscopy has been hindered by the complexity and cost of its implementation. Gated CW-STED (gCW-STED) substantially helps solve this problem without sacrificing spatial resolution. Here, we describe a versatile gCW-STED microscope able to speedily image the specimen, at a resolution below 50nm, with light intensities comparable to the more complicated all-pulsed STED system. We show this ability on calibration samples as well as on biological samples.
ISSN:1046-2023
1095-9130
DOI:10.1016/j.ymeth.2013.06.029