Employing Flow Cytometry to Extracellular Vesicles Sample Microvolume Analysis and Quality Control

Area of Science:

• Biotechnology
• Cell Biology
• Biomarker Discovery

Background:

• Extracellular Vesicles (EVs) mediate cell-cell communication and are valuable non-invasive biomarkers in liquid biopsies.
• Current EV analysis often requires large sample volumes for isolation, limiting applications.
• Efficient EV analysis methods are needed for small or challenging biofluid samples.

Purpose of the Study:

• To develop a novel flow cytometry (FC) strategy for direct EV analysis without prior isolation or concentration.
• To enable population analysis of EVs in limited sample volumes and reduce processing time.
• To provide a tool for quality control of EV isolation protocols.

Main Methods:

• A direct flow cytometry (FC) staining and analysis strategy for extracellular vesicles (EVs).
• Application in longitudinal analysis of EVs in mouse plasma and murine vitreous humor.
• Quantification of vesicular particles in purified and non-purified biological samples.

Main Results:

• Demonstrated successful population analysis of EVs in small sample volumes (mouse plasma, vitreous humor).
• Reduced sample processing time compared to traditional isolation methods.
• Provided a method for quality control of EV isolates from different purification protocols.

Conclusions:

• The developed FC strategy allows direct EV analysis, overcoming sample volume limitations.
• This method enhances the clinical potential of liquid biopsies for biomarker discovery.
• It offers a versatile tool for EV research and quality assessment.