Details

Autor(en) / Beteiligte

• Kaufmann, Rainer
• Schellenberger, Pascale
• Seiradake, Elena
• Dobbie, Ian M
• Jones, E. Yvonne
• Davis, Ilan
• Hagen, Christoph
• Grünewald, Kay

Titel

Super-Resolution Microscopy Using Standard Fluorescent Proteins in Intact Cells under Cryo-Conditions

Ist Teil von

• Nano letters, 2014-07, Vol.14 (7), p.4171-4175

Ort / Verlag

Washington, DC: American Chemical Society

Erscheinungsjahr

2014

Link zum Volltext

Quelle

MEDLINE

Beschreibungen/Notizen

• We introduce a super-resolution technique for fluorescence cryo-microscopy based on photoswitching of standard genetically encoded fluorescent marker proteins in intact mammalian cells at low temperature (81 K). Given the limit imposed by the lack of cryo-immersion objectives, current applications of fluorescence cryo-microscopy to biological specimens achieve resolutions between 400–500 nm only. We demonstrate that the single molecule characteristics of reversible photobleaching of mEGFP and mVenus at liquid nitrogen temperature are suitable for the basic concept of single molecule localization microscopy. This enabled us to perform super-resolution imaging of vitrified biological samples and to visualize structures in unperturbed fast frozen cells for the first time with a structural resolution of ∼125 nm (average single molecule localization accuracy ∼40 nm), corresponding to a 3–5 fold resolution improvement.