STED with wavelengths closer to the emission maximum

In stimulated emission depletion (STED) nanoscopy the wavelength of the STED beam is usually tuned towards the red tail of the emission maximum of the fluorophore. Shifting the STED wavelength closer to the emission peak, i.e. towards the blue region, favorably increases the stimulated emission cros...

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Hauptverfasser: Vicidomini, Giuseppe (VerfasserIn) , Hell, Stefan (VerfasserIn)
Dokumenttyp: Article (Journal)
Sprache:Englisch
Veröffentlicht: 16 February 2012
In: Optics express
Year: 2012, Jahrgang: 20, Heft: 5, Pages: 5225-5236
ISSN:1094-4087
DOI:10.1364/OE.20.005225
Online-Zugang:Verlag, Volltext: http://dx.doi.org/10.1364/OE.20.005225
Verlag, Volltext: https://www.osapublishing.org/oe/abstract.cfm?uri=oe-20-5-5225
Volltext
Verfasserangaben:Giuseppe Vicidomini, Gael Moneron, Christian Eggeling, Eva Rittweger, and Stefan W. Hell

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520 |a In stimulated emission depletion (STED) nanoscopy the wavelength of the STED beam is usually tuned towards the red tail of the emission maximum of the fluorophore. Shifting the STED wavelength closer to the emission peak, i.e. towards the blue region, favorably increases the stimulated emission cross-section. However, this blue-shifting also increases the probability to excite fluorophores that have remained in their ground state, compromising the image contrast. Here we present a method to exploit the higher STED efficiency of blue-shifted STED beams while maintaining the contrast in the image. The method is exemplified by imaging immunolabeled features in mammalian cells with an up to 3-fold increased STED efficiency compared to that encountered in standard STED nanoscopy implementations. 
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