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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...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
2016
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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. |
format | Online Article Text |
id | pubmed-5014615 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT daimingjie opticalvisualisationofindividualbiomoleculesindenselypackedclusters AT jungmannralf opticalvisualisationofindividualbiomoleculesindenselypackedclusters AT yinpeng opticalvisualisationofindividualbiomoleculesindenselypackedclusters |