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Rapid and accurate analysis of stem cell-derived extracellular vesicles with super resolution microscopy and live imaging

Extracellular vesicles (EVs) have prevalent roles in cancer biology and regenerative medicine. Conventional techniques for characterising EVs including electron microscopy (EM), nanoparticle tracking analysis (NTA) and tuneable resistive pulse sensing (TRPS), have been reported to produce high varia...

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Autores principales: Nizamudeen, Zubair, Markus, Robert, Lodge, Rhys, Parmenter, Christopher, Platt, Mark, Chakrabarti, Lisa, Sottile, Virginie
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6203808/
https://www.ncbi.nlm.nih.gov/pubmed/30290236
http://dx.doi.org/10.1016/j.bbamcr.2018.09.008
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author Nizamudeen, Zubair
Markus, Robert
Lodge, Rhys
Parmenter, Christopher
Platt, Mark
Chakrabarti, Lisa
Sottile, Virginie
author_facet Nizamudeen, Zubair
Markus, Robert
Lodge, Rhys
Parmenter, Christopher
Platt, Mark
Chakrabarti, Lisa
Sottile, Virginie
author_sort Nizamudeen, Zubair
collection PubMed
description Extracellular vesicles (EVs) have prevalent roles in cancer biology and regenerative medicine. Conventional techniques for characterising EVs including electron microscopy (EM), nanoparticle tracking analysis (NTA) and tuneable resistive pulse sensing (TRPS), have been reported to produce high variability in particle count (EM) and poor sensitivity in detecting EVs below 50 nm in size (NTA and TRPS), making accurate and unbiased EV analysis technically challenging. This study introduces direct stochastic optical reconstruction microscopy (d-STORM) as an efficient and reliable characterisation approach for stem cell-derived EVs. Using a photo-switchable lipid dye, d-STORM imaging enabled rapid detection of EVs down to 20–30 nm in size with higher sensitivity and lower variability compared to EM, NTA and TRPS techniques. Imaging of EV uptake by live stem cells in culture further confirmed the potential of this approach for downstream cell biology applications and for the analysis of vesicle-based cell-cell communication.
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spelling pubmed-62038082018-12-01 Rapid and accurate analysis of stem cell-derived extracellular vesicles with super resolution microscopy and live imaging Nizamudeen, Zubair Markus, Robert Lodge, Rhys Parmenter, Christopher Platt, Mark Chakrabarti, Lisa Sottile, Virginie Biochim Biophys Acta Mol Cell Res Article Extracellular vesicles (EVs) have prevalent roles in cancer biology and regenerative medicine. Conventional techniques for characterising EVs including electron microscopy (EM), nanoparticle tracking analysis (NTA) and tuneable resistive pulse sensing (TRPS), have been reported to produce high variability in particle count (EM) and poor sensitivity in detecting EVs below 50 nm in size (NTA and TRPS), making accurate and unbiased EV analysis technically challenging. This study introduces direct stochastic optical reconstruction microscopy (d-STORM) as an efficient and reliable characterisation approach for stem cell-derived EVs. Using a photo-switchable lipid dye, d-STORM imaging enabled rapid detection of EVs down to 20–30 nm in size with higher sensitivity and lower variability compared to EM, NTA and TRPS techniques. Imaging of EV uptake by live stem cells in culture further confirmed the potential of this approach for downstream cell biology applications and for the analysis of vesicle-based cell-cell communication. Elsevier 2018-12 /pmc/articles/PMC6203808/ /pubmed/30290236 http://dx.doi.org/10.1016/j.bbamcr.2018.09.008 Text en © 2018 The Author(s) http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Nizamudeen, Zubair
Markus, Robert
Lodge, Rhys
Parmenter, Christopher
Platt, Mark
Chakrabarti, Lisa
Sottile, Virginie
Rapid and accurate analysis of stem cell-derived extracellular vesicles with super resolution microscopy and live imaging
title Rapid and accurate analysis of stem cell-derived extracellular vesicles with super resolution microscopy and live imaging
title_full Rapid and accurate analysis of stem cell-derived extracellular vesicles with super resolution microscopy and live imaging
title_fullStr Rapid and accurate analysis of stem cell-derived extracellular vesicles with super resolution microscopy and live imaging
title_full_unstemmed Rapid and accurate analysis of stem cell-derived extracellular vesicles with super resolution microscopy and live imaging
title_short Rapid and accurate analysis of stem cell-derived extracellular vesicles with super resolution microscopy and live imaging
title_sort rapid and accurate analysis of stem cell-derived extracellular vesicles with super resolution microscopy and live imaging
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6203808/
https://www.ncbi.nlm.nih.gov/pubmed/30290236
http://dx.doi.org/10.1016/j.bbamcr.2018.09.008
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