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Deussner-Helfmann, Nina S.; Auer, Alexander; Strauss, Maximilian T.; Malkusch, Sebastian; Dietz, Marina S.; Barth, Hans -Dieter; Jungmann, Ralf; Heilemann, Mike (2018): Correlative Single-Molecule FRET and DNA-PAINT Imaging. In: Nano Letters, Vol. 18, No. 7: pp. 4626-4630
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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 hither extract FRET values from single binding sites that are spaced just similar to 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.