Extracellular Vesicles and Fluorescence Excitation-Emission Matrix Spectroscopy

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Extracellular vesicles (EVs) are emerging as key players in intercellular communication, acting as messengers that shuttle proteins, lipids, and nucleic acids between cells. In bodily fluids such as blood, urine, saliva, and breast milk, EVs are gaining significant interest in normal physiology and disease states. Fluorescence spectroscopy, a powerful analytical tool, offers unique advantages for characterizing EVs, aiding researchers in unlocking the secrets of cellular communication.

EVs have captured the scientific spotlight in recent years. These tiny membranous structures, released by various cell types, have emerged as essential cellular communication components. EVs ferry critical biological cargo, including proteins, lipids, and nucleic acids, between cells, influencing a range of physiological processes. While the initial exploration of EVs began in the 1980s, the field has witnessed an exponential surge in interest and research activity over the past two decades.

Understanding the role of EVs in health and disease necessitates effective methods for their analysis. EV analysis encompasses a toolbox of techniques crucial for characterizing their size, concentration, and cargo composition. Additionally, pinpointing the cell of origin and elucidating how recipient cells take up EVs and respond to their cargo are critical aspects of EV research.

Fluorescence spectroscopy allows researchers to non-destructively probe the biomolecules carried within EVs based on their unique spectroscopic properties. Fluorescence spectroscopy offers several advantages, including high sensitivity, specificity, and the ability to perform label-free analysis, preserving the integrity of the EVs. By analyzing the fluorescence signature of EVs, researchers can gain insights into the types of molecules they contain, providing valuable clues about their potential functions.

EVs, including exosomes, microvesicles, and apoptotic bodies, are small, membrane-bound particles released by cells into the extracellular environment. These vesicles are involved in numerous physiological processes, including immune responses, cell signaling, and the transfer of genetic material. EVs have gained attention as potential biomarkers for various diseases, such as cancer, neurodegenerative disorders, and cardiovascular diseases, due to their cargo (i.e., proteins, lipids, and nucleic acids) (1).

As mentioned earlier, the characterization of EVs is crucial for understanding their biological functions and potential therapeutic applications. Traditional methods, such as nanoparticle tracking analysis (NTA) and electron microscopy, provide valuable information about EV size and morphology. Fluorescence spectroscopy, a technique that measures the emission of light by fluorescent molecules within a sample, can offer a rapid and sensitive complementary alternative technique for EV characterization.

This study aims to investigate the application of fluorescence spectroscopy for the analysis of EVs derived from different cell types. Fluorescence could be another powerful complementary technique in EV research’s toolbox.

Materials and methods

Materials and reagents

Lyophilized human embryonic kidney 293 (HEK293) cell-derived exosomes were obtained from Hansa BioMed (Tallinn, Estonia). Additionally, lyophilized exosome standards, PC-3 (ab239689) and MCF7 (ab239691), each at a concentration of 1 x 1012, were purchased from Abcam (Cambridge, Mass.). All samples were stored at -80 °C. In addition, 1X phosphate-buffered saline (PBS) was obtained from Corning (Corning, NY).

Read the article in BioPharm International’s Manufacturing and Facilities eBook.

Reference

1. Yarlott, L.K.; Gilmore, A. M. A-TEEM Spectroscopic Characterization of Exosome Standards and Their Mixtures. Poster presentation at the International Society for Extracellular Vesicles (ISEV) Annual Meeting, Virtual, May 18–21, 2021.

About the author

Lyufei Chen, PhD, is an application scientist at HORIBA Scientific.

Article details

BioPharm International®
eBook: Manufacturing and Facilities
November 2024
Pages: 22–26

Citation

When referring to this article, please cite it as Chen, L. Extracellular Vesicles and Fluorescence Excitation-Emission Matrix Spectroscopy. BioPharm International Manufacturing and Facilities eBook, November 2024.