Correlative Single-Molecule FRET and DNA-PAINT Imaging.
Nano Lett
; 18(7): 4626-4630, 2018 07 11.
Article
em En
| MEDLINE
| ID: mdl-29943993
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 â¼55 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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Base de dados:
MEDLINE
Assunto principal:
DNA
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Nanotecnologia
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Transferência Ressonante de Energia de Fluorescência
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Imagem Individual de Molécula
Idioma:
En
Ano de publicação:
2018
Tipo de documento:
Article