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Dual-color DNA-PAINT single-particle tracking enables extended studies of membrane protein interactions

DNA-PAINT based single-particle tracking (DNA-PAINT-SPT) has recently significantly enhanced observation times in in vitro SPT experiments by overcoming the constraints of fluorophore photobleaching. However, with the reported implementation, only a single target can be imaged and the technique cann...

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Autores principales: Niederauer, Christian, Nguyen, Chikim, Wang-Henderson, Miles, Stein, Johannes, Strauss, Sebastian, Cumberworth, Alexander, Stehr, Florian, Jungmann, Ralf, Schwille, Petra, Ganzinger, Kristina A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10356854/
https://www.ncbi.nlm.nih.gov/pubmed/37468504
http://dx.doi.org/10.1038/s41467-023-40065-8
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author Niederauer, Christian
Nguyen, Chikim
Wang-Henderson, Miles
Stein, Johannes
Strauss, Sebastian
Cumberworth, Alexander
Stehr, Florian
Jungmann, Ralf
Schwille, Petra
Ganzinger, Kristina A.
author_facet Niederauer, Christian
Nguyen, Chikim
Wang-Henderson, Miles
Stein, Johannes
Strauss, Sebastian
Cumberworth, Alexander
Stehr, Florian
Jungmann, Ralf
Schwille, Petra
Ganzinger, Kristina A.
author_sort Niederauer, Christian
collection PubMed
description DNA-PAINT based single-particle tracking (DNA-PAINT-SPT) has recently significantly enhanced observation times in in vitro SPT experiments by overcoming the constraints of fluorophore photobleaching. However, with the reported implementation, only a single target can be imaged and the technique cannot be applied straight to live cell imaging. Here we report on leveraging this technique from a proof-of-principle implementation to a useful tool for the SPT community by introducing simultaneous live cell dual-color DNA-PAINT-SPT for quantifying protein dimerization and tracking proteins in living cell membranes, demonstrating its improved performance over single-dye SPT.
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spelling pubmed-103568542023-07-21 Dual-color DNA-PAINT single-particle tracking enables extended studies of membrane protein interactions Niederauer, Christian Nguyen, Chikim Wang-Henderson, Miles Stein, Johannes Strauss, Sebastian Cumberworth, Alexander Stehr, Florian Jungmann, Ralf Schwille, Petra Ganzinger, Kristina A. Nat Commun Article DNA-PAINT based single-particle tracking (DNA-PAINT-SPT) has recently significantly enhanced observation times in in vitro SPT experiments by overcoming the constraints of fluorophore photobleaching. However, with the reported implementation, only a single target can be imaged and the technique cannot be applied straight to live cell imaging. Here we report on leveraging this technique from a proof-of-principle implementation to a useful tool for the SPT community by introducing simultaneous live cell dual-color DNA-PAINT-SPT for quantifying protein dimerization and tracking proteins in living cell membranes, demonstrating its improved performance over single-dye SPT. Nature Publishing Group UK 2023-07-19 /pmc/articles/PMC10356854/ /pubmed/37468504 http://dx.doi.org/10.1038/s41467-023-40065-8 Text en © The Author(s) 2023 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Niederauer, Christian
Nguyen, Chikim
Wang-Henderson, Miles
Stein, Johannes
Strauss, Sebastian
Cumberworth, Alexander
Stehr, Florian
Jungmann, Ralf
Schwille, Petra
Ganzinger, Kristina A.
Dual-color DNA-PAINT single-particle tracking enables extended studies of membrane protein interactions
title Dual-color DNA-PAINT single-particle tracking enables extended studies of membrane protein interactions
title_full Dual-color DNA-PAINT single-particle tracking enables extended studies of membrane protein interactions
title_fullStr Dual-color DNA-PAINT single-particle tracking enables extended studies of membrane protein interactions
title_full_unstemmed Dual-color DNA-PAINT single-particle tracking enables extended studies of membrane protein interactions
title_short Dual-color DNA-PAINT single-particle tracking enables extended studies of membrane protein interactions
title_sort dual-color dna-paint single-particle tracking enables extended studies of membrane protein interactions
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10356854/
https://www.ncbi.nlm.nih.gov/pubmed/37468504
http://dx.doi.org/10.1038/s41467-023-40065-8
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