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Correlative Single-Molecule FRET and DNA-PAINT Imaging.
- Source :
-
Nano letters [Nano Lett] 2018 Jul 11; Vol. 18 (7), pp. 4626-4630. Date of Electronic Publication: 2018 Jun 29. - Publication Year :
- 2018
-
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.
Details
- Language :
- English
- ISSN :
- 1530-6992
- Volume :
- 18
- Issue :
- 7
- Database :
- MEDLINE
- Journal :
- Nano letters
- Publication Type :
- Academic Journal
- Accession number :
- 29943993
- Full Text :
- https://doi.org/10.1021/acs.nanolett.8b02185