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Optical visualisation of individual biomolecules in densely packed clusters

Recent advances in fluorescence super-resolution microscopy have allowed sub-cellular features and synthetic nanostructures down to ~15 nm in size to be imaged. However, direct optical observation of individual molecular targets (~5 nm) in a densely packed biomolecular cluster remains a challenge. H...

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Detalles Bibliográficos
Autores principales: Dai, Mingjie, Jungmann, Ralf, Yin, Peng
Formato: Online Artículo Texto
Lenguaje:English
Publicado: 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5014615/
https://www.ncbi.nlm.nih.gov/pubmed/27376244
http://dx.doi.org/10.1038/nnano.2016.95
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author Dai, Mingjie
Jungmann, Ralf
Yin, Peng
author_facet Dai, Mingjie
Jungmann, Ralf
Yin, Peng
author_sort Dai, Mingjie
collection PubMed
description Recent advances in fluorescence super-resolution microscopy have allowed sub-cellular features and synthetic nanostructures down to ~15 nm in size to be imaged. However, 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 hybridisation - on synthetic DNA nanostructures. We examined the effects of high photon count, high blinking statistics, and appropriate blinking duty cycle on imaging quality, and developed a software-based drift correction method that achieves <1 nm residual drift (r.m.s.) over hours. This allowed us to image a densely packed triangular lattice pattern with ~5 nm point-to-point distance, and analyse DNA origami structural offset with angstrom-level precision (2 Å) from single-molecule studies. By combining the approach with multiplexed Exchange-PAINT imaging, we further demonstrated an optical nano-display with 5×5 nm pixel size and three distinct colours, and with <1 nm cross-channel registration accuracy. This method opens up possibilities for direct and quantitative optical observation of individual biomolecular features in crowded environments.
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spelling pubmed-50146152017-01-04 Optical visualisation of individual biomolecules in densely packed clusters Dai, Mingjie Jungmann, Ralf Yin, Peng Nat Nanotechnol Article Recent advances in fluorescence super-resolution microscopy have allowed sub-cellular features and synthetic nanostructures down to ~15 nm in size to be imaged. However, 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 hybridisation - on synthetic DNA nanostructures. We examined the effects of high photon count, high blinking statistics, and appropriate blinking duty cycle on imaging quality, and developed a software-based drift correction method that achieves <1 nm residual drift (r.m.s.) over hours. This allowed us to image a densely packed triangular lattice pattern with ~5 nm point-to-point distance, and analyse DNA origami structural offset with angstrom-level precision (2 Å) from single-molecule studies. By combining the approach with multiplexed Exchange-PAINT imaging, we further demonstrated an optical nano-display with 5×5 nm pixel size and three distinct colours, and with <1 nm cross-channel registration accuracy. This method opens up possibilities for direct and quantitative optical observation of individual biomolecular features in crowded environments. 2016-07-04 2016-09 /pmc/articles/PMC5014615/ /pubmed/27376244 http://dx.doi.org/10.1038/nnano.2016.95 Text en Users may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use:http://www.nature.com/authors/editorial_policies/license.html#terms
spellingShingle Article
Dai, Mingjie
Jungmann, Ralf
Yin, Peng
Optical visualisation of individual biomolecules in densely packed clusters
title Optical visualisation of individual biomolecules in densely packed clusters
title_full Optical visualisation of individual biomolecules in densely packed clusters
title_fullStr Optical visualisation of individual biomolecules in densely packed clusters
title_full_unstemmed Optical visualisation of individual biomolecules in densely packed clusters
title_short Optical visualisation of individual biomolecules in densely packed clusters
title_sort optical visualisation of individual biomolecules in densely packed clusters
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5014615/
https://www.ncbi.nlm.nih.gov/pubmed/27376244
http://dx.doi.org/10.1038/nnano.2016.95
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