Site‐specific labeling of Affimers for DNA‐PAINT microscopy

Abstract

Optical super‐resolution techniques allow fluorescence imaging below the classical diffraction limit of light. From a technology standpoint of view, recent methods are approaching molecular‐scale spatial resolution. However, this remarkable achievement is not easily translated to imaging of cellular components, as 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). Here, we combine recently developed Affimer reagents with site‐specific DNA modification for high‐efficiency labeling and imaging using DNA‐PAINT. We assay our approach using an actin Affimer. The small DNA‐conjugated affinity binders could provide a solution for efficient multi‐target super‐resolution imaging in the future.

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Item Type:	Article
Authors/Creators:	Schlichthaerle, T Eklund, A Schueder, F Strauss, M Tiede, C https://orcid.org/0000-0003-0280-4005 Curd, A https://orcid.org/0000-0002-3949-7523 Ries, J Peckham, M https://orcid.org/0000-0002-3754-2028 Tomlinson, D https://orcid.org/0000-0003-4134-7484 Jungmann, R
Copyright, Publisher and Additional Information:	© 2018 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim. This is the peer reviewed version of the following article: Schlichthaerle, T. , Eklund, A. , Schueder, F. , Strauss, M. , Tiede, C. , Curd, A. , Ries, J. , Peckham, M. , Tomlinson, D. and Jungmann, R. (2018), Site‐specific labeling of Affimers for DNA‐PAINT microscopy. Angew. Chem. Int. Ed.. Accepted Author Manuscript. . doi:10.1002/anie.201804020, which has been published in final form at https://doi.org/10.1002/anie.201804020. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving. Uploaded in accordance with the publisher's self-archiving policy.
Keywords:	Small Binders; super-resolution microscopy; DNA-PAINT; Quantitative Microscopy
Dates:	Published: 20 August 2018 Published (online): 6 June 2018 Accepted: 5 June 2018
Institution:	The University of Leeds
Academic Units:	The University of Leeds > Faculty of Biological Sciences (Leeds) > School of Molecular and Cellular Biology (Leeds) > Cell Biology (Leeds) The University of Leeds > Faculty of Biological Sciences (Leeds) > School of Molecular and Cellular Biology (Leeds) > Synthetic Biology (Leed)
Funding Information:	Funder Grant number Wellcome Trust 098249/Z/12/Z MRC MR/K015613/1
Depositing User:	Symplectic Publications
Date Deposited:	12 Jun 2018 10:16
Last Modified:	06 Jun 2019 00:42
Status:	Published
Publisher:	Wiley
Identification Number:	10.1002/anie.201804020
Open Archives Initiative ID (OAI ID):	oai:eprints.whiterose.ac.uk:131878

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Site‐specific labeling of Affimers for DNA‐PAINT microscopy

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