Open AccessPitfalls associated with lipophilic fluorophore staining of extracellular vesicles for uptake studies
Author(s) -
Simonsen Jens B.
Publication year - 2019
Publication title -
journal of extracellular vesicles
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.94
H-Index - 68
ISSN - 2001-3078
DOI - 10.1080/20013078.2019.1582237
Subject(s) - extracellular vesicles , vesicle , fluorophore , staining , extracellular vesicle , amphiphile , microvesicles , extracellular , biophysics , microbiology and biotechnology , chemistry , biochemistry , biology , fluorescence , membrane , copolymer , genetics , microrna , physics , organic chemistry , quantum mechanics , gene , polymer
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.