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Monomeric Garnet, a far-red fluorescent protein for live-cell STED imaging

Hense, A.; Prunsche, B.; Gao, P.; Ishitsuka, Y.; Nienhaus, K.; Nienhaus, G.U.

The advancement of far-red emitting variants of the green fluorescent protein (GFP) is crucially important for imaging live cells, tissues and organisms. Despite notable efforts, far-red marker proteins still need further optimization to match the performance of their green counterparts. Here we present mGarnet, a robust monomeric marker protein with far-red fluorescence peaking at 670 nm. Thanks to its large extinction coefficient of 95,000 M-1 cm-1, mGarnet can be efficiently excited with 640-nm light on the red edge of its 598-nm excitation band. A large Stokes shift allows essentially the entire fluorescence emission to be collected even with 640-nm excitation, counterbalancing the lower fluorescence quantum yield of mGarnet, 9.1%, that is typical of far-red FPs. We demonstrate an excellent performance as a live-cell fusion marker in STED microscopy, using 640 nm excitation and 780 nm depletion wavelengths.

Volltext §
DOI: 10.5445/IR/1000052538
DOI: 10.1038/srep18006
Zitationen: 26
Web of Science
Zitationen: 24
Zitationen: 35
Cover der Publikation
Zugehörige Institution(en) am KIT Institut für Angewandte Physik (APH)
Institut für Nanotechnologie (INT)
Institut für Toxikologie und Genetik (ITG)
Karlsruhe School of Optics & Photonics (KSOP)
Publikationstyp Zeitschriftenaufsatz
Publikationsjahr 2015
Sprache Englisch
Identifikator ISSN: 2045-2322
KITopen-ID: 1000052538
HGF-Programm 43.23.01 (POF III, LK 01) Advanced Optical Lithography+Microscopy
Erschienen in Scientific Reports
Verlag Nature Research
Band 5
Seiten 18006/1-10
Bemerkung zur Veröffentlichung Gefördert durch den KIT-Publikationsfonds
Nachgewiesen in Web of Science
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