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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 ...

 

 

 

 

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

Publication:
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