Conventional, High-Resolution and Imaging Flow Cytometry: Benchmarking Performance in Characterisation of...

Area of Science:

• Biomedical Engineering
• Analytical Chemistry
• Cell Biology

Background:

• Flow cytometry is vital for high-throughput particle analysis.
• Conventional flow cytometry has limitations in extracellular vesicle (EV) sensitivity.
• Novel high-resolution and imaging flow cytometry platforms offer enhanced EV detection capabilities.

Purpose of the Study:

• To comparatively benchmark conventional, high-resolution, and imaging flow cytometry platforms for EV characterization.
• To evaluate the sensitivity, quantification accuracy, and repeatability of different flow cytometry platforms for EV analysis.
• To guide researchers in selecting appropriate flow cytometry platforms for EV studies.

Main Methods:

• Comparative benchmarking of FACS Aria III, Apogee A60 Micro-PLUS, and ImageStream X Mk II flow cytometers.
• Characterization of nanospheres with varying sizes, refractive indices, and fluorescence properties.
• Assessment of EV detection and concentration variability in blood plasma across platforms.

Main Results:

• Nanospheres were used to systematically characterize platform abilities in detecting and quantifying particle populations.
• Repeatability in concentration determinations was reported for different nanosphere populations.
• Variabilities in EV concentration determination (intra-day, inter-day, global) were evaluated for the three platforms.

Conclusions:

• Comparative analysis provides benchmarks for selecting suitable flow cytometry platforms for EV research.
• Informed platform selection enhances the robustness and reproducibility of extracellular vesicle studies.
• This methodology aids researchers in matching project needs with specific flow cytometry capabilities.