Published on March 18, 2026
Double-emulsion (DE) droplets enable isolation of single cells together with assay reagents, allowing sorting and recovery of selected cells based on functional readouts. Unlike conventional water-in-oil droplets, DE droplets consist of an aqueous core surrounded by an oil shell and suspended in an outer aqueous phase. This structure makes them directly compatible with conventional fluorescence-activated cell sorting (FACS) systems, enabling integration of droplet microfluidics and flow cytometry.
For Xdrop workflows, this compatibility is essential for selective recovery of fluorescently labelled droplets containing cells of interest.
Xdrop DE50 droplets have a diameter of approximately 75 µm, making them substantially larger than typical mammalian cells (10-20 µm). In addition, the oil shell contributes to increased mass and altered hydrodynamic behavior.
When standard cell-sorting parameters are applied without adjustment, users may see:
Sorting double-emulsion droplets requires optimization of fluidics and timing parameters to accommodate their size and hydrodynamic behavior. Adjustments to nozzle configuration, pressure settings, and drop delay are central to maintaining droplet integrity and sort precision.
To address these considerations, Sony Biotechnology developed a Large Particle Sorting software option for the Sony MA900 and Sony SH800 cell sorters. This software is specifically designed to optimize instrument performance when handling large and heavy particles such as double-emulsion droplets.
Key adaptations include:
Optimized droplet breakoff: Adjusted fluidic control to stabilize droplet formation under modified pressure and nozzle conditions appropriate for larger events.
Refined timing and drop-delay settings: Parameter adjustments for the altered velocity and mass of large particles, improving accurate deposition into collection tubes.
Prioritized gating: The option to prioritize sorting thereby limiting the number of aborted events and boosting recovery.
For laboratories using Sony platforms, these features simplify implementation of droplet-based sorting workflows and enhance reproducibility.
Double-emulsion droplets expand the capabilities of functional single-cell analysis by enabling high-throughput screening with downstream recovery of selected cells. However, the mechanical and hydrodynamic challenges associated with sorting large DE droplets require sort parameter optimization.
Sony’s Large Particle Sorting software illustrates how instrument-level adaptations can support reliable sorting of DE droplets. When combined with droplet microfluidics workflows such as Xdrop, these optimizations enable efficient, high-precision recovery of functionally selected single cells.
Figure: Combined workflow between Xdrop and Sony MA900 to identify cell-cell interactions
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Flow cytometry analysis and sorting of double-emulsion droplets protocol
Sorting DE50 droplets with Large Particle Sorting Option - Sony MA900 and SH800 cell sorters protocol
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