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REVIEW article
Front. Bioeng. Biotechnol.
Sec. Nanobiotechnology
Volume 12 - 2024 |
doi: 10.3389/fbioe.2024.1479516
Factors to consider before choosing EV labeling method for fluorescence-based techniques
Provisionally accepted- Chair and Department of Biochemistry, Medical University of Warsaw, Warsaw, Poland
A well-designed fluorescence-based analysis of extracellular vesicles (EV) can provide insights into the size, morphology, and biological function of EVs, which can be used in medical applications. Fluorescent nanoparticle tracking analysis with appropriate controls can provide reliable data for size and concentration measurements, while nanoscale flow cytometry is the most appropriate tool for characterizing molecular cargoes. Label selection is a crucial element in all fluorescence methods. The most comprehensive data can be obtained if several labeling approaches for a given marker are used, as they would provide complementary information about EV populations and interactions with the cells. In all EV-related experiments, the influence of lipoproteins and protein corona on the results should be considered. By reviewing and considering all the factors affecting EV labeling methods used in fluorescence-based techniques, we can assert that the data will provide as accurate as possible information about true EV biology and offer precise, clinically applicable information for future EVbased diagnostic or therapeutic applications.
Keywords: extracellular vesicles, f-NTA, nanoscale flow cytometry, Fluorescent staining, corona, Lipophilic dyes, Lipoproteins
Received: 12 Aug 2024; Accepted: 05 Sep 2024.
Copyright: © 2024 Dlugolecka and Czystowska-Kuzmicz. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence:
Malgorzata Czystowska-Kuzmicz, Chair and Department of Biochemistry, Medical University of Warsaw, Warsaw, Poland
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