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

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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Veröffentlicht in:Journal of extracellular vesicles 2019-12, Vol.8 (1), p.1582237-n/a
1. Verfasser: Simonsen, Jens B.
Format: Artikel
Sprache:eng
Schlagworte:
Biomarkers
Chromatography
contamination
dissociation
Extracellular vesicles
Fluorophores
High density lipoprotein
Labeling
labelling
Lipids
Lipophilic
Lipoproteins
Nanoparticles
Plasma
Proteins
staining
Stains & staining
targeting
tracking
uptake
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Zusammenfassung: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.
ISSN:2001-3078
2001-3078
DOI:10.1080/20013078.2019.1582237