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Nat Nanotechnol. 2016 Sep;11(9):798-807. doi: 10.1038/nnano.2016.95. Epub 2016 Jul 4.

Optical imaging of individual biomolecules in densely packed clusters.

Dai M1,2, Jungmann R1,3, Yin P1,3.

Author information

1
Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts 02115, USA.
2
Biophysics Program, Harvard University, Boston, Massachusetts 02115, USA.
3
Department of Systems Biology, Harvard Medical School, Boston, Massachusetts 02115, USA.

Abstract

Recent advances in fluorescence super-resolution microscopy have allowed subcellular features and synthetic nanostructures down to 10-20 nm in size to be imaged. However, the direct optical observation of individual molecular targets (∼5 nm) in a densely packed biomolecular cluster remains a challenge. Here, we show that such discrete molecular imaging is possible using DNA-PAINT (points accumulation for imaging in nanoscale topography)-a super-resolution fluorescence microscopy technique that exploits programmable transient oligonucleotide hybridization-on synthetic DNA nanostructures. We examined the effects of a high photon count, high blinking statistics and an appropriate blinking duty cycle on imaging quality, and developed a software-based drift correction method that achieves <1 nm residual drift (root mean squared) over hours. This allowed us to image a densely packed triangular lattice pattern with ∼5 nm point-to-point distance and to analyse the DNA origami structural offset with ångström-level precision (2 Å) from single-molecule studies. By combining the approach with multiplexed exchange-PAINT imaging, we further demonstrated an optical nanodisplay with 5 × 5 nm pixel size and three distinct colours with <1 nm cross-channel registration accuracy.

PMID:
27376244
PMCID:
PMC5014615
DOI:
10.1038/nnano.2016.95
[Indexed for MEDLINE]
Free PMC Article

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