Quantum-yield-optimized fluorophores for site-specific labeling and super-resolution imaging.
J. Am. Chem. Soc.. 2011-06-01; 133(21): 8090-8093
DOI: 10.1021/ja200967z
Read on PubMed
1. J Am Chem Soc. 2011 Jun 1;133(21):8090-3. doi: 10.1021/ja200967z. Epub 2011 May
11.
Quantum-yield-optimized fluorophores for site-specific labeling and
super-resolution imaging.
Grunwald C(1), Schulze K, Giannone G, Cognet L, Lounis B, Choquet D, Tampé R.
Author information:
(1)Institute of Biochemistry, Goethe-University Frankfurt, Max-von-Laue-Str. 9,
D-60438 Frankfurt/M., Germany.
Single-molecule applications, saturated pattern excitation microscopy, and
stimulated emission depletion (STED) microscopy demand bright as well as highly
stable fluorescent dyes. Here we describe the synthesis of
quantum-yield-optimized fluorophores for reversible, site-specific labeling of
proteins or macromolecular complexes. We used polyproline-II (PPII) helices as
sufficiently rigid spacers with various lengths to improve the fluorescence
signals of a set of different trisNTA-fluorophores. The improved quantum yields
were demonstrated by steady-state and fluorescence lifetime analyses. As a proof
of principle, we characterized the trisNTA-PPII-fluorophores with respect to in
vivo protein labeling and super-resolution imaging at synapses of living neurons.
The distribution of His-tagged AMPA receptors (GluA1) in spatially restricted
synaptic clefts was imaged by confocal and STED microscopy. The comparison of
fluorescence intensity profiles revealed the superior resolution of STED
microscopy. These results highlight the advantages of biocompatible and, in
particular, small and photostable trisNTA-PPII-fluorophores in super-resolution
microscopy.
© 2011 American Chemical Society
DOI: 10.1021/ja200967z
PMID: 21545135 [Indexed for MEDLINE]