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. 2018 Jul 11;18(7):4626-4630.
doi: 10.1021/acs.nanolett.8b02185. Epub 2018 Jun 29.

Correlative Single-Molecule FRET and DNA-PAINT Imaging

Nina S Deußner-Helfmann  1 , Alexander Auer  2   3 , Maximilian T Strauss  2   3 , Sebastian Malkusch  1 , Marina S Dietz  1 , Hans-Dieter Barth  1 , Ralf Jungmann  2   3 , Mike Heilemann  1
Affiliations

Correlative Single-Molecule FRET and DNA-PAINT Imaging

Nina S Deußner-Helfmann et al. Nano Lett. .

Abstract

DNA-PAINT is an optical super-resolution microscopy method that can visualize nanoscale protein arrangements and provide spectrally unlimited multiplexing capabilities. However, current multiplexing implementations based on, for example, DNA exchange (such as Exchange-PAINT) achieves multitarget detection by sequential imaging, limiting throughput. Here, we combine DNA-PAINT with single-molecule FRET and use the FRET efficiency as parameter for multiplexed imaging with high specificity. We demonstrate correlated single-molecule FRET and super-resolution on DNA origami structures, which are equipped with binding sequences that are targeted by pairs of dye-labeled oligonucleotides generating the FRET signal. We futher extract FRET values from single binding sites that are spaced just ∼55 nm apart, demonstrating super-resolution FRET imaging. This combination of FRET and DNA-PAINT allows for multiplexed super-resolution imaging with low background and opens the door for accurate distance readout in the 1-10 nm range.

Keywords: DNA origami; DNA-PAINT; Single-molecule FRET; multiplexing; super-resolution microscopy.

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