Journal
NANO LETTERS
Volume 18, Issue 7, Pages 4626-4630Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.nanolett.8b02185
Keywords
Single-molecule FRET; DNA-PAINT; super-resolution microscopy; DNA origami; multiplexing
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Funding
- DFG [SFB 902]
- DFG through the Graduate School of Quantitative Biosciences Munich (QBM)
- International Max Planck Research School for Molecular and Cellular Life Sciences (IMPRS-LS)
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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.
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