Open Access Original article

Novel red fluorophores with superior performance in STED microscopy

Christian A Wurm12*, Kirill Kolmakov1, Fabian Göttfert1, Haisen Ta1, Mariano Bossi3, Heiko Schill1, Sebastian Berning1, Stefan Jakobs12, Gerald Donnert1, Vladimir N Belov1* and Stefan W Hell1*

Author Affiliations

1 Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, Göttingen, 37077, Germany

2 Department of Neurology, Georg-August University of Göttingen Medical School, Göttingen, 37073, Germany

3 Inquimae Conicet-UBA, Pabellon 2, Ciudad Universitaria, Buenos Aires, Argentina

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Optical Nanoscopy 2012, 1:7  doi:10.1186/2192-2853-1-7

Published: 28 September 2012

Abstract

In optical microscopy, most red-emitting dyes provide only moderate performance due to unspecific binding, poor labeling efficiency, and insufficient brightness. Here we report on four novel red fluororescent dyes, including the first phosphorylated dye, created by combining a rigidized rhodamine backbone with various polar groups. They exhibit large fluorescence quantum yields and improved NHS ester stability. While these fluorophores are highly suitable for fluorescence microscopy in general, they excel in stimulated emission depletion (STED) microscopy, providing < 25 nm spatial resolution in raw images of cells.

Keywords:
Microscopy; Confocal; STED; Fluorescence; Rhodamines; Bioconjugation; Super-resolution; Nanoscopy; Stimulated emission depletion