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Fluorescence lifetime DNA-PAINT for multiplexed super-resolution imaging of cells
DNA point accumulation for imaging in nanoscale topography (DNA-PAINT) is a powerful super-resolution technique highly suitable for multi-target (multiplexing) bio-imaging. However, multiplexed imaging of cells is still challenging due to the dense and sticky environment inside a cell. Here, we comb...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8752799/ https://www.ncbi.nlm.nih.gov/pubmed/35017652 http://dx.doi.org/10.1038/s42003-021-02976-4 |
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author | Oleksiievets, Nazar Sargsyan, Yelena Thiele, Jan Christoph Mougios, Nikolaos Sograte-Idrissi, Shama Nevskyi, Oleksii Gregor, Ingo Opazo, Felipe Thoms, Sven Enderlein, Jörg Tsukanov, Roman |
author_facet | Oleksiievets, Nazar Sargsyan, Yelena Thiele, Jan Christoph Mougios, Nikolaos Sograte-Idrissi, Shama Nevskyi, Oleksii Gregor, Ingo Opazo, Felipe Thoms, Sven Enderlein, Jörg Tsukanov, Roman |
author_sort | Oleksiievets, Nazar |
collection | PubMed |
description | DNA point accumulation for imaging in nanoscale topography (DNA-PAINT) is a powerful super-resolution technique highly suitable for multi-target (multiplexing) bio-imaging. However, multiplexed imaging of cells is still challenging due to the dense and sticky environment inside a cell. Here, we combine fluorescence lifetime imaging microscopy (FLIM) with DNA-PAINT and use the lifetime information as a multiplexing parameter for targets identification. In contrast to Exchange-PAINT, fluorescence lifetime PAINT (FL-PAINT) can image multiple targets simultaneously and does not require any fluid exchange, thus leaving the sample undisturbed and making the use of flow chambers/microfluidic systems unnecessary. We demonstrate the potential of FL-PAINT by simultaneous imaging of up to three targets in a cell using both wide-field FLIM and 3D time-resolved confocal laser scanning microscopy (CLSM). FL-PAINT can be readily combined with other existing techniques of multiplexed imaging and is therefore a perfect candidate for high-throughput multi-target bio-imaging. |
format | Online Article Text |
id | pubmed-8752799 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-87527992022-01-20 Fluorescence lifetime DNA-PAINT for multiplexed super-resolution imaging of cells Oleksiievets, Nazar Sargsyan, Yelena Thiele, Jan Christoph Mougios, Nikolaos Sograte-Idrissi, Shama Nevskyi, Oleksii Gregor, Ingo Opazo, Felipe Thoms, Sven Enderlein, Jörg Tsukanov, Roman Commun Biol Article DNA point accumulation for imaging in nanoscale topography (DNA-PAINT) is a powerful super-resolution technique highly suitable for multi-target (multiplexing) bio-imaging. However, multiplexed imaging of cells is still challenging due to the dense and sticky environment inside a cell. Here, we combine fluorescence lifetime imaging microscopy (FLIM) with DNA-PAINT and use the lifetime information as a multiplexing parameter for targets identification. In contrast to Exchange-PAINT, fluorescence lifetime PAINT (FL-PAINT) can image multiple targets simultaneously and does not require any fluid exchange, thus leaving the sample undisturbed and making the use of flow chambers/microfluidic systems unnecessary. We demonstrate the potential of FL-PAINT by simultaneous imaging of up to three targets in a cell using both wide-field FLIM and 3D time-resolved confocal laser scanning microscopy (CLSM). FL-PAINT can be readily combined with other existing techniques of multiplexed imaging and is therefore a perfect candidate for high-throughput multi-target bio-imaging. Nature Publishing Group UK 2022-01-11 /pmc/articles/PMC8752799/ /pubmed/35017652 http://dx.doi.org/10.1038/s42003-021-02976-4 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Oleksiievets, Nazar Sargsyan, Yelena Thiele, Jan Christoph Mougios, Nikolaos Sograte-Idrissi, Shama Nevskyi, Oleksii Gregor, Ingo Opazo, Felipe Thoms, Sven Enderlein, Jörg Tsukanov, Roman Fluorescence lifetime DNA-PAINT for multiplexed super-resolution imaging of cells |
title | Fluorescence lifetime DNA-PAINT for multiplexed super-resolution imaging of cells |
title_full | Fluorescence lifetime DNA-PAINT for multiplexed super-resolution imaging of cells |
title_fullStr | Fluorescence lifetime DNA-PAINT for multiplexed super-resolution imaging of cells |
title_full_unstemmed | Fluorescence lifetime DNA-PAINT for multiplexed super-resolution imaging of cells |
title_short | Fluorescence lifetime DNA-PAINT for multiplexed super-resolution imaging of cells |
title_sort | fluorescence lifetime dna-paint for multiplexed super-resolution imaging of cells |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8752799/ https://www.ncbi.nlm.nih.gov/pubmed/35017652 http://dx.doi.org/10.1038/s42003-021-02976-4 |
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