Pitfalls associated with lipophilic fluorophore staining of extracellular vesicles for uptake studies

Post-staining of extracellular vesicles (EVs) with lipid-anchored fluorophores (LAFs) such as PKH67 is a widely used strategy for studying EVs but it is associated with several pitfalls. The pitfalls discussed in this commentary are related to LAF labelling of non-EV species due to (1) lipoprotein c...

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Autor principal: Simonsen, Jens B.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Taylor & Francis 2019
Materias:
Technical Report
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6383605/
https://www.ncbi.nlm.nih.gov/pubmed/30815239
http://dx.doi.org/10.1080/20013078.2019.1582237
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author Simonsen, Jens B.
author_facet Simonsen, Jens B.
author_sort Simonsen, Jens B.
collection PubMed
description Post-staining of extracellular vesicles (EVs) with lipid-anchored fluorophores (LAFs) such as PKH67 is a widely used strategy for studying EVs but it is associated with several pitfalls. The pitfalls discussed in this commentary are related to LAF labelling of non-EV species due to (1) lipoprotein contamination in EV samples, (2) desorption of the LAF reporters from vesicles into proteins and lipoproteins in blood and serum, and (3) the capability of the amphiphilic LAF compounds to form EV-like particles. Awareness of these challenges and developing solutions to overcome these are important to ensure that we make relevant interpretations when using LAFs to track EVs.
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spelling pubmed-63836052019-02-27 Pitfalls associated with lipophilic fluorophore staining of extracellular vesicles for uptake studies Simonsen, Jens B. J Extracell Vesicles Technical Report Post-staining of extracellular vesicles (EVs) with lipid-anchored fluorophores (LAFs) such as PKH67 is a widely used strategy for studying EVs but it is associated with several pitfalls. The pitfalls discussed in this commentary are related to LAF labelling of non-EV species due to (1) lipoprotein contamination in EV samples, (2) desorption of the LAF reporters from vesicles into proteins and lipoproteins in blood and serum, and (3) the capability of the amphiphilic LAF compounds to form EV-like particles. Awareness of these challenges and developing solutions to overcome these are important to ensure that we make relevant interpretations when using LAFs to track EVs. Taylor & Francis 2019-02-20 /pmc/articles/PMC6383605/ /pubmed/30815239 http://dx.doi.org/10.1080/20013078.2019.1582237 Text en © 2019 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group on behalf of The International Society for Extracellular Vesicles. http://creativecommons.org/licenses/by-nc/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Technical Report
Simonsen, Jens B.
Pitfalls associated with lipophilic fluorophore staining of extracellular vesicles for uptake studies
title Pitfalls associated with lipophilic fluorophore staining of extracellular vesicles for uptake studies
title_full Pitfalls associated with lipophilic fluorophore staining of extracellular vesicles for uptake studies
title_fullStr Pitfalls associated with lipophilic fluorophore staining of extracellular vesicles for uptake studies
title_full_unstemmed Pitfalls associated with lipophilic fluorophore staining of extracellular vesicles for uptake studies
title_short Pitfalls associated with lipophilic fluorophore staining of extracellular vesicles for uptake studies
title_sort pitfalls associated with lipophilic fluorophore staining of extracellular vesicles for uptake studies
topic Technical Report
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6383605/
https://www.ncbi.nlm.nih.gov/pubmed/30815239
http://dx.doi.org/10.1080/20013078.2019.1582237
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