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PLoS One. 2012;7(6):e40003. doi: 10.1371/journal.pone.0040003. Epub 2012 Jun 27.

Achieving λ/10 resolution CW STED nanoscopy with a Ti:Sapphire oscillator.

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1
School of Biomedical Engineering, Shanghai Jiao Tong University, Shanghai, China.

Abstract

In this report, a Ti:Sapphire oscillator was utilized to realize synchronization-free stimulated emission depletion (STED) microscopy. With pump power of 4.6 W and sample irradiance of 310 mW, we achieved super-resolution as high as 71 nm. With synchronization-free STED, we imaged 200 nm nanospheres as well as all three cytoskeletal elements (microtubules, intermediate filaments, and actin filaments), clearly demonstrating the resolving power of synchronization-free STED over conventional diffraction limited imaging. It also allowed us to discover that, Dylight 650, exhibits improved performance over ATTO647N, a fluorophore frequently used in STED. Furthermore, we applied synchronization-free STED to image fluorescently-labeled intracellular viral RNA granules, which otherwise cannot be differentiated by confocal microscopy. Thanks to the widely available Ti:Sapphire oscillators in multiphoton imaging system, this work suggests easier access to setup super-resolution microscope via the synchronization-free STED.

PMID:
22761944
PMCID:
PMC3384643
DOI:
10.1371/journal.pone.0040003
[Indexed for MEDLINE]
Free PMC Article
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