Live‐cell super‐resolution imaging of actin using LifeAct‐14 with a PAINT‐based approach

We present direct‐LIVE‐PAINT, an easy‐to‐implement approach for the nanoscopic imaging of protein structures in live cells using labeled binding peptides. We demonstrate the feasibility of direct‐LIVE‐PAINT with an actin‐binding peptide fused to EGFP, the location of which can be accurately determin...

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Detalles Bibliográficos
Autores principales: Bhaskar, Haresh, Kleinjan, Dirk‐Jan, Oi, Curran, Gidden, Zoe, Rosser, Susan J., Horrocks, Mathew H., Regan, Lynne
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley & Sons, Inc. 2023
Materias:
For the Record
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9878614/
https://www.ncbi.nlm.nih.gov/pubmed/36585831
http://dx.doi.org/10.1002/pro.4558
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author Bhaskar, Haresh
Kleinjan, Dirk‐Jan
Oi, Curran
Gidden, Zoe
Rosser, Susan J.
Horrocks, Mathew H.
Regan, Lynne
author_facet Bhaskar, Haresh
Kleinjan, Dirk‐Jan
Oi, Curran
Gidden, Zoe
Rosser, Susan J.
Horrocks, Mathew H.
Regan, Lynne
author_sort Bhaskar, Haresh
collection PubMed
description We present direct‐LIVE‐PAINT, an easy‐to‐implement approach for the nanoscopic imaging of protein structures in live cells using labeled binding peptides. We demonstrate the feasibility of direct‐LIVE‐PAINT with an actin‐binding peptide fused to EGFP, the location of which can be accurately determined as it transiently binds to actin filaments. We show that direct‐LIVE‐PAINT can be used to image actin structures below the diffraction‐limit of light and have used it to observe the dynamic nature of actin in live cells. We envisage a similar approach could be applied to imaging other proteins within live mammalian cells.
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spelling pubmed-98786142023-02-01 Live‐cell super‐resolution imaging of actin using LifeAct‐14 with a PAINT‐based approach Bhaskar, Haresh Kleinjan, Dirk‐Jan Oi, Curran Gidden, Zoe Rosser, Susan J. Horrocks, Mathew H. Regan, Lynne Protein Sci For the Record We present direct‐LIVE‐PAINT, an easy‐to‐implement approach for the nanoscopic imaging of protein structures in live cells using labeled binding peptides. We demonstrate the feasibility of direct‐LIVE‐PAINT with an actin‐binding peptide fused to EGFP, the location of which can be accurately determined as it transiently binds to actin filaments. We show that direct‐LIVE‐PAINT can be used to image actin structures below the diffraction‐limit of light and have used it to observe the dynamic nature of actin in live cells. We envisage a similar approach could be applied to imaging other proteins within live mammalian cells. John Wiley & Sons, Inc. 2023-02-01 /pmc/articles/PMC9878614/ /pubmed/36585831 http://dx.doi.org/10.1002/pro.4558 Text en © 2022 The Authors. Protein Science published by Wiley Periodicals LLC on behalf of The Protein Society. https://creativecommons.org/licenses/by/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle For the Record
Bhaskar, Haresh
Kleinjan, Dirk‐Jan
Oi, Curran
Gidden, Zoe
Rosser, Susan J.
Horrocks, Mathew H.
Regan, Lynne
Live‐cell super‐resolution imaging of actin using LifeAct‐14 with a PAINT‐based approach
title Live‐cell super‐resolution imaging of actin using LifeAct‐14 with a PAINT‐based approach
title_full Live‐cell super‐resolution imaging of actin using LifeAct‐14 with a PAINT‐based approach
title_fullStr Live‐cell super‐resolution imaging of actin using LifeAct‐14 with a PAINT‐based approach
title_full_unstemmed Live‐cell super‐resolution imaging of actin using LifeAct‐14 with a PAINT‐based approach
title_short Live‐cell super‐resolution imaging of actin using LifeAct‐14 with a PAINT‐based approach
title_sort live‐cell super‐resolution imaging of actin using lifeact‐14 with a paint‐based approach
topic For the Record
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9878614/
https://www.ncbi.nlm.nih.gov/pubmed/36585831
http://dx.doi.org/10.1002/pro.4558
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