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Correlative single-molecule FRET and DNA-PAINT imaging

July 2018. 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. A team of scientists from Goethe University Frankfurt and the Max Planck Institute of Biochemistry have now combined DNA-PAINT with single-molecule FRET and use the FRET efficiency as parameter for multiplexed imaging with high specificity. The scientists demonstrate in their new publication in the journal Nano Letters 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. They also extracted 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.More ...





Mike Heilemann, Institute of Physical and Theoretical Chemistry, Goethe University Frankfurt, Riedberg Campus, 60438 Frankfurt/Main, Germany, Heilemann@chemie.uni-frankfurt.de

Nina Deußner-Helfmann, Alexander Auer, Maximilian Strauss, Sebastian Malkusch, Marina Dietz, Hans-Dieter Barth, Ralf Jungmann* and Mike Heilemann* (2018) Correlative single-molecule FRET and DNA-PAINT imaging. Nano Letters: published online 26 June 2018. dx.doi.org/10.1021/acs.nanolett.8b02185