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

David Virant, Bjoern Traenkle, Julia Maier, Philipp D. Kaiser, Mona Bodenhöfer, Christian Schmees, Ilijana Vojnovic, Borbála Pisak-Lukáts, Ulrike Endesfelder () and Ulrich Rothbauer ()
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David Virant: Max Planck Institute for Terrestrial Microbiology and LOEWE Center for Synthetic Microbiology (SYNMIKRO)
Bjoern Traenkle: Eberhard Karls University Tuebingen
Julia Maier: Eberhard Karls University Tuebingen
Philipp D. Kaiser: Natural and Medical Sciences Institute at the University of Tuebingen
Mona Bodenhöfer: Natural and Medical Sciences Institute at the University of Tuebingen
Christian Schmees: Natural and Medical Sciences Institute at the University of Tuebingen
Ilijana Vojnovic: Max Planck Institute for Terrestrial Microbiology and LOEWE Center for Synthetic Microbiology (SYNMIKRO)
Borbála Pisak-Lukáts: Max Planck Institute for Terrestrial Microbiology and LOEWE Center for Synthetic Microbiology (SYNMIKRO)
Ulrike Endesfelder: Max Planck Institute for Terrestrial Microbiology and LOEWE Center for Synthetic Microbiology (SYNMIKRO)
Ulrich Rothbauer: Eberhard Karls University Tuebingen

Nature Communications, 2018, vol. 9, issue 1, 1-14

Abstract: 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.

Date: 2018
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DOI: 10.1038/s41467-018-03191-2

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