An extensive step-by-step protocol describing the workflow is available in the Supporting Information . Picoliter DEs were generated using four syringe pumps (PicoPump Elite, Harvard Apparatus) for cell suspension and inner, oil, and outer sheath solutions. The inner phase was composed of 1× PBS with 0.5% BSA. The oil phase was composed of HFE7500 fluorinated oil (Sigma) and 2.2% Ionic PEG-Krytox34 (link) (FSH, Miller-Stephenson). The carrier phase contained 1% Tween-20 (Sigma) and 2% Pluronic F68 (Kolliphor 188, Sigma) in PBS.31 (link) Each phase was loaded into syringes (PlastiPak, BD) and connected to the device via PE/2 tubing (Scientific Commodities). Relatively low flow rates (400:125:105:6000 μL/h, oil.cell/reagent/outer) were used to reduce cell shear stress.
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Picoliter Droplet Generation for Cell Encapsulation
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Corresponding Organization : Chan Zuckerberg Biohub San Francisco
Other organizations : California Institute for Regenerative Medicine
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Variable analysis
independent variables
- Flow rates of the oil, cell/reagent, and outer phases
- Not explicitly mentioned
- Composition of the inner phase (1× PBS with 0.5% BSA)
- Composition of the oil phase (HFE7500 fluorinated oil and 2.2% Ionic PEG-Krytox)
- Composition of the carrier phase (1% Tween-20 and 2% Pluronic F68 in PBS)
- No positive or negative controls were explicitly mentioned in the provided information.
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