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Angew Chem Int Ed Engl. 2018 Aug 20;57(34):11060-11063. doi: 10.1002/anie.201804020. Epub 2018 Jul 15.

Site-Specific Labeling of Affimers for DNA-PAINT Microscopy.

Author information

1
Faculty of Physics and Center for Nanoscience, LMU Munich, Munich, Germany.
2
Max Planck Institute of Biochemistry, Martinsried, Germany.
3
Astbury Centre for Structural and Molecular Biology, University of Leeds, Leeds, UK.
4
School of Molecular and Cellular Biology, University of Leeds, Leeds, UK.
5
European Molecular Biology Laboratory, Cell Biology and Biophysics Unit, Heidelberg, Germany.

Abstract

Optical super-resolution techniques allow fluorescence imaging below the classical diffraction limit of light. From a technology standpoint, recent methods are approaching molecular-scale spatial resolution. However, this remarkable achievement is not easily translated to imaging of cellular components, since current labeling approaches are limited by either large label sizes (antibodies) or the sparse availability of small and efficient binders (nanobodies, aptamers, genetically-encoded tags). In this work, we combined recently developed Affimer reagents with site-specific DNA modification for high-efficiency labeling and imaging using DNA-PAINT. We assayed our approach using an actin Affimer. The small DNA-conjugated affinity binders could provide a solution for efficient multitarget super-resolution imaging in the future.

KEYWORDS:

Affimers; DNA-PAINT; quantitative microscopy; small binders; super-resolution microscopy

PMID:
29873161
DOI:
10.1002/anie.201804020
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