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Deterministic droplet-based co-encapsulation of single cells through inertial and hydrodynamic focusing
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DOI：10.1039/D5AN00986C.png)
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Abstract
En 中文
Microfluidic techniques for high-throughput encapsulation are powerful tools in single-cell analytics and cytokine profiling. Inertial focusing microfluidics is widely used to align particles in uniform sequences; enhancing encapsulation efficiency. However; on-chip sample dilution strategies to further optimize efficiency remain largely unexplored in deterministic encapsulation approaches; both experimentally and through theoretical modeling. Here; we present a high-yield microparticle encapsulation method that combines inertial and hydrodynamic focusing to enable precise tuning of microparticle spacing and modulation of capture efficiency; thereby offering enhanced operational flexibility for controlled particle encapsulation. We first investigate the microparticle self-ordering behavior within the spiral loop and characterize both flow dynamics and droplet formation regimes. By varying the sheath-to-sample flow rate ratio from 0 to 2; we observe that higher ratios increase the interparticle spacing and shift particles closer to the channel wall. These trends align with both analytical modeling and 3D numerical simulations. Notably; at higher sheath flow ratios (e.g.; 1 and 2); single-particle encapsulation exceeds 76%; significantly surpassing Poisson distribution predictions. Moreover; single-cell capture efficiency exceeds 60% under these conditions. In co-encapsulation experiments; we achieved a one-cell-multiple-beads co-encapsulation efficiency near 40%; marking a significant improvement over the Poisson limit. For single-cell applications; we performed co-encapsulation of THP-1 monocytes and streptavidin-coated magnetic beads for TNF-α cytokine detection following lipopolysaccharide stimulation. Cytokine secretion was successfully detected at the single-cell level in both aqueous droplets and alginate hydrogels. We anticipate that this method will offer a promising platform for probing cell–cell interactions and immune responses at single-cell resolution.
Keywords:
microfluidics
single-cell encapsulation
inertial focusing
hydrodynamic focusing
droplet-based co-encapsulation
Journal
A
IF:
3.3
Papers:
1.6W
Citations:
3.3W