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Proceedings Paper

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

Correlating DNA-PAINT (point accumulation for imaging in nanoscale topography) and single-molecule FRET (Förster 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.

Paper Details

Date Published: 13 February 2020
PDF: 7 pages
Proc. SPIE 11246, Single Molecule Spectroscopy and Superresolution Imaging XIII, 112460L (13 February 2020); doi: 10.1117/12.2550629
Show Author Affiliations
Nina S. Deussner-Helfmann, Goethe-Univ. Frankfurt am Main (Germany)
Alexander Auer, Ludwig-Maximilians-Univ. München (Germany)
Max-Planck-Institut für Biochemie (Germany)
Maximilian T. Strauss, Ludwig-Maximilians-Univ. München (Germany)
Max-Planck-Institut für Biochemie (Germany)
Paul Donlin-Asp, Max Planck Institute of Brain Research (Germany)
Sebastian Malkusch, Goethe-Univ. Frankfurt am Main (Germany)
Marina S. Dietz, Goethe-Univ. Frankfurt am Main (Germany)
Hans-Dieter Barth, Goethe-Univ. Frankfurt am Main (Germany)
Erin Schuman, Max Planck Institute of Brain Research (Germany)
Ralf Jungmann, Ludwig-Maximilians-Univ. München (Germany)
Max-Planck-Institut für Biochemie (Germany)
Mike Heilemann, Goethe-Univ. Frankfurt am Main (Germany)


Published in SPIE Proceedings Vol. 11246:
Single Molecule Spectroscopy and Superresolution Imaging XIII
Ingo Gregor; Felix Koberling; Rainer Erdmann, Editor(s)

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