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Updated: Jan 22, 2026
09:30
Purification and Analysis of Caenorhabditis elegans Extracellular Vesicles
Published on: March 31, 2020
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FCMPASS Software Aids Extracellular Vesicle Light Scatter Standardization.
Joshua A Welsh1,2, Peter Horak3, James S Wilkinson3
1Faculty of Medicine, University of Southampton, Southampton, UK.
Summary
Standardizing flow cytometry for extracellular vesicle (EV) analysis is crucial. This study introduces FCMPASS software to standardize light scatter measurements, improving EV characterization and reproducibility in research.
Area of Science:
- Biotechnology
- Analytical Chemistry
- Nanomedicine
Background:
- Extracellular vesicles (EVs) show significant promise in clinical applications like diagnostics and therapeutics.
- Flow cytometry is a widely used, high-throughput method for EV analysis, but lacks standardization.
- Current flow cytometers often operate at their detection limits for EVs, hindering accurate analysis.
Purpose of the Study:
- To develop and demonstrate a method for standardizing light scatter measurements in flow cytometry for EV analysis.
- To introduce flow cytometer post-acquisition analysis software (FCMPASS) for improved EV characterization.
Main Methods:
- Utilized Mie theory within the FCMPASS software for flow cytometer light scatter standardization.
- Employed beads of known diameter and refractive index, or known collection angles, to approximate geometric parameters.
- Generated scatter-diameter and scatter-refractive index curves for data standardization.
Main Results:
- FCMPASS enables the conversion of scattering data and instrument sensitivity into standardized units.
- The software allows for the conversion of light scatter data into diameter or refractive index distributions with appropriate controls.
- Demonstrated a method to standardize flow cytometer geometric parameters.
Conclusions:
- FCMPASS provides a freely available and user-friendly approach to standardize EV characterization using flow cytometry.
- Standardization enhances the interpretability, comparability, and reproducibility of EV flow cytometry research.
- This method extends the capabilities of flow cytometry for detailed EV analysis.
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