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Nat Commun. 2018 Mar 2;9(1):930. doi: 10.1038/s41467-018-03191-2.

A peptide tag-specific nanobody enables high-quality labeling for dSTORM imaging.

Author information

1
Department of Systems and Synthetic Microbiology, Max Planck Institute for Terrestrial Microbiology and LOEWE Center for Synthetic Microbiology (SYNMIKRO), Karl-von-Frisch Strasse 16, Marburg, 35043, Germany.
2
Pharmaceutical Biotechnology, Eberhard Karls University Tuebingen, Markwiesenstrasse 55, Reutlingen, 72770, Germany.
3
Natural and Medical Sciences Institute at the University of Tuebingen, Markwiesenstrasse 55, Reutlingen, 72770, Germany.
4
Department of Systems and Synthetic Microbiology, Max Planck Institute for Terrestrial Microbiology and LOEWE Center for Synthetic Microbiology (SYNMIKRO), Karl-von-Frisch Strasse 16, Marburg, 35043, Germany. ulrike.endesfelder@synmikro.mpi-marburg.mpg.de.
5
Pharmaceutical Biotechnology, Eberhard Karls University Tuebingen, Markwiesenstrasse 55, Reutlingen, 72770, Germany. ulrich.rothbauer@uni-tuebingen.de.
6
Natural and Medical Sciences Institute at the University of Tuebingen, Markwiesenstrasse 55, Reutlingen, 72770, Germany. ulrich.rothbauer@uni-tuebingen.de.

Abstract

Dense fluorophore labeling without compromising the biological target is crucial for genuine super-resolution microscopy. Here we introduce a broadly applicable labeling strategy for fixed and living cells utilizing a short peptide tag-specific nanobody (BC2-tag/bivBC2-Nb). BC2-tagging of ectopically introduced or endogenous proteins does not interfere with the examined structures and bivBC2-Nb staining results in a close-grained fluorophore labeling with minimal linkage errors. This allowed us to perform high-quality dSTORM imaging of various targets in mammalian and yeast cells. We expect that this versatile strategy will render many more demanding cellular targets amenable to dSTORM imaging.

PMID:
29500346
PMCID:
PMC5834503
DOI:
10.1038/s41467-018-03191-2
[Indexed for MEDLINE]
Free PMC Article

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