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  Correlating DNA-PAINT and single-molecule FRET for multiplexed super-resolution imaging

Deussner-Helfmann, N. S., Auer, A., Strauss, M. T., Donlin-Asp, P., Malkusch, S., Dietz, M. S., et al. (2020). Correlating DNA-PAINT and single-molecule FRET for multiplexed super-resolution imaging. Proceedings of SPIE, 11246: 112460L. doi:10.1117/12.2550629.

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 Creators:
Deussner-Helfmann, Nina S.1, Author
Auer, Alexander2, Author              
Strauss, Maximilian T.2, Author              
Donlin-Asp, Paul1, Author
Malkusch, Sebastian1, Author
Dietz, Marina S.1, Author
Barth, Hans-Dieter1, Author
Schuman, Erin1, Author
Jungmann, Ralf2, Author              
Heilemann, Mike1, Author
Affiliations:
1external, ou_persistent22              
2Jungmann, Ralf / Molecular Imaging and Bionanotechnology, Max Planck Institute of Biochemistry, Max Planck Society, ou_2149679              

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Free keywords: LOCALIZATION MICROSCOPY; RESOLUTION; BINDINGSingle-molecule FRET; DNA-PAINT; super-resolution microscopy; DNA origami; multiplexing;
 Abstract: Correlating DNA-PAINT (point accumulation for imaging in nanoscale topography) and single-molecule FRET (Forster resonance energy transfer) enables the multiplexed detection with sub-diffraction optical resolution. We designed pairs of short oligonucleotides, labeled with donor and acceptor fluorophores with various distances generating different FRET efficiencies. The strands can transiently bind to a target docking strand, simultaneous binding of both strands results in FRET signals which yield a super-resolved image via DNA-PAINT imaging. We demonstrate FRET-PAINT by designing and imaging DNA origami, which is a useful tool to establish super-resolution methods. The DNA origami structures were equipped with three target binding sites spaced by 55 nm, a sub-diffraction limited distance, however ensuring that no FRET between the target sites occurs. We resolved the individual binding sites in the donor and acceptor channels, and in addition extracted the FRET efficiency for each site in single and mixed populations. The combination of FRET and DNA-PAINT allows for multiplexed super-resolution imaging in conjunction with distance-sensitive readout in the 1 to 10 nm range.

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Language(s): eng - English
 Dates: 2020
 Publication Status: Published online
 Pages: 7
 Publishing info: -
 Table of Contents: -
 Rev. Type: -
 Identifiers: ISI: 000546225400007
DOI: 10.1117/12.2550629
 Degree: -

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Title: SPIE BiOS
Place of Event: San Francisco, CA
Start-/End Date: 2020-02-01 - 2020-02-06

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Title: Proceedings of SPIE
  Other : Single Molecule Spectroscopy and Superresolution Imaging XIII
Source Genre: Journal
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Publ. Info: Bellingham, Washington : SPIE
Pages: - Volume / Issue: 11246 Sequence Number: 112460L Start / End Page: - Identifier: ISSN: 0277-786X
CoNE: https://pure.mpg.de/cone/journals/resource/0277-786X