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Optimization of Flow Cytometric Sorting Parameters for High-Throughput Isolation and Purification of Small Extracellular Vesicles
Published on: January 20, 2023
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Enhancing large particle recovery in high-throughput functional cell sorting through ΔBOP optimization.
Naohisa Sakamoto1, Eikichi Shibata1,2, Mitsuo Yoshimura3
1Life Science Technology Research & Development Department, Technology Development Laboratories, Sony Corporation, Tokyo, Japan.
Scientific Reports
|December 21, 2025
Summary
Sorting large particles in flow cytometry is difficult. This study introduces a novel ΔBOP strategy to improve droplet stability and significantly increase recovery rates for single-cell analysis.
Area of Science:
- Biotechnology
- Biophysics
- Analytical Chemistry
Background:
- Flow cytometry is essential for single-cell analysis.
- Sorting large particles (>30 μm) presents recovery challenges.
Purpose of the Study:
- Investigate particle size impact on Break-off point (BOP).
- Assess large particle effects on droplet formation.
- Develop a strategy to enhance recovery rates.
Main Methods:
- Combined simulations and experiments.
- Analyzed particle-induced and vibration-driven BOP.
- Introduced ΔBOP (difference between BOPs) for optimization.
Main Results:
- Droplet instability due to BOP interference limits recovery.
- ΔBOP optimization stabilized droplet formation.
- Achieved 92.4% recovery for 35-µm and 75.6% for 50-µm particles.
Conclusions:
- The ΔBOP strategy enhances recovery for large particles.
- Applicable to various particle types and high-throughput analyses.
- Enables new research in secretion profiling and intercellular communication.
