MRT letter: Nanoscopy of protein colocalization in living cells by STED and GSDIM

We report the use of superresolution fluorescence microscopy for studying the nanoscale distribution of protein colocalization in living mammalian cells. Nanoscale imaging is attained both by a targeted and a stochastic fluorescence on-off switching superresolution method, namely by stimulated emiss...

Ausführliche Beschreibung

Gespeichert in:
Bibliographische Detailangaben
Hauptverfasser: Lalkens, Birka (VerfasserIn) , Hell, Stefan (VerfasserIn)
Dokumenttyp: Article (Journal)
Sprache:Englisch
Veröffentlicht: 2012
In: Microscopy research and technique
Year: 2012, Jahrgang: 75, Heft: 1, Pages: 1-6
ISSN:1097-0029
DOI:10.1002/jemt.21026
Online-Zugang:Verlag, Volltext: http://dx.doi.org/10.1002/jemt.21026
Verlag, Volltext: https://onlinelibrary.wiley.com/doi/abs/10.1002/jemt.21026
Volltext
Verfasserangaben:Birka Lalkens, Ilaria Testa, Katrin I. Willig, and Stefan W. Hell
Beschreibung
Zusammenfassung:We report the use of superresolution fluorescence microscopy for studying the nanoscale distribution of protein colocalization in living mammalian cells. Nanoscale imaging is attained both by a targeted and a stochastic fluorescence on-off switching superresolution method, namely by stimulated emission depletion (STED) and ground state depletion microscopy followed by individual molecular return (GSDIM), respectively. Analysis of protein colocalization is performed by bimolecular fluorescence complementation (BiFC). Specifically, a nonfluorescent fragment of the yellow fluorescent protein Citrine is fused to tubulin while a counterpart nonfluorescent fragment is fused to the microtubulin-associated protein MAP2 such that fluorescence is reconstituted on contact of the fragment-carrying proteins. Images with resolution down to 65 nm prove a powerful new way for studying protein colocalization in living cells at the nanoscale. Microsc. Res. Tech., 2012. © 2011 Wiley Periodicals, Inc.
Beschreibung:Published online 15 June 2011
Gesehen am 31.10.2018
Beschreibung:Online Resource
ISSN:1097-0029
DOI:10.1002/jemt.21026