Real-time fluorescence imaging with 20 nm axial resolution

Measuring the nanoscale organization of protein structures near the plasma membrane of live cells is challenging, especially when the structure is dynamic. Here we present the development of a two-wavelength total internal reflection fluorescence method capable of real-time imaging of cellular struc...

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Autores principales: Stabley, Daniel R., Oh, Thomas, Simon, Sanford M., Mattheyses, Alexa L., Salaita, Khalid
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Pub. Group 2015
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4595625/
https://www.ncbi.nlm.nih.gov/pubmed/26392382
http://dx.doi.org/10.1038/ncomms9307
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author Stabley, Daniel R.
Oh, Thomas
Simon, Sanford M.
Mattheyses, Alexa L.
Salaita, Khalid
author_facet Stabley, Daniel R.
Oh, Thomas
Simon, Sanford M.
Mattheyses, Alexa L.
Salaita, Khalid
author_sort Stabley, Daniel R.
collection PubMed
description Measuring the nanoscale organization of protein structures near the plasma membrane of live cells is challenging, especially when the structure is dynamic. Here we present the development of a two-wavelength total internal reflection fluorescence method capable of real-time imaging of cellular structure height with nanometre resolution. The method employs a protein of interest tagged with two different fluorophores and imaged to obtain the ratio of emission in the two channels. We use this approach to visualize the nanoscale organization of microtubules and endocytosis of the epidermal growth factor receptor.
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spelling pubmed-45956252015-10-21 Real-time fluorescence imaging with 20 nm axial resolution Stabley, Daniel R. Oh, Thomas Simon, Sanford M. Mattheyses, Alexa L. Salaita, Khalid Nat Commun Article Measuring the nanoscale organization of protein structures near the plasma membrane of live cells is challenging, especially when the structure is dynamic. Here we present the development of a two-wavelength total internal reflection fluorescence method capable of real-time imaging of cellular structure height with nanometre resolution. The method employs a protein of interest tagged with two different fluorophores and imaged to obtain the ratio of emission in the two channels. We use this approach to visualize the nanoscale organization of microtubules and endocytosis of the epidermal growth factor receptor. Nature Pub. Group 2015-09-22 /pmc/articles/PMC4595625/ /pubmed/26392382 http://dx.doi.org/10.1038/ncomms9307 Text en Copyright © 2015, Nature Publishing Group, a division of Macmillan Publishers Limited. All Rights Reserved. http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Stabley, Daniel R.
Oh, Thomas
Simon, Sanford M.
Mattheyses, Alexa L.
Salaita, Khalid
Real-time fluorescence imaging with 20 nm axial resolution
title Real-time fluorescence imaging with 20 nm axial resolution
title_full Real-time fluorescence imaging with 20 nm axial resolution
title_fullStr Real-time fluorescence imaging with 20 nm axial resolution
title_full_unstemmed Real-time fluorescence imaging with 20 nm axial resolution
title_short Real-time fluorescence imaging with 20 nm axial resolution
title_sort real-time fluorescence imaging with 20 nm axial resolution
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4595625/
https://www.ncbi.nlm.nih.gov/pubmed/26392382
http://dx.doi.org/10.1038/ncomms9307
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AT salaitakhalid realtimefluorescenceimagingwith20nmaxialresolution

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