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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...

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Autores principales: Oleksiievets, Nazar, Sargsyan, Yelena, Thiele, Jan Christoph, Mougios, Nikolaos, Sograte-Idrissi, Shama, Nevskyi, Oleksii, Gregor, Ingo, Opazo, Felipe, Thoms, Sven, Enderlein, Jörg, Tsukanov, Roman
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2022
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.
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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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