Rigid 20–40 μm beads made from hydroxylated methacrylic polymer (Toyopearl HW-65S, Tosoh Bioscience) are used to demonstrate the formation of monodispersed bead-containing droplets at limiting dilution. Beads are suspended in 68% OptiPrep DensityGradient Medium (Sigma-Aldrich), and co-flowed with 2% agarose (Ultra-low Gelling Temperature Agarose, Type IX-A, Sigma-Aldrich) in 1× PBS. The device, identical to one previously described,1 (link) is fabricated at a height of 100 μm. Flow rates are 2000 μl h–1 bead-containing solution, 2000 μl h–1 agarose, and 40 000 μl h–1 2% ionic Krytox, prepared as previously described.18 (link)
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Monodispersed Bead-Containing Droplet Formation
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Agarose
Device
Dilution
Ionic
Low temperature
Polymer
Rigid
Corresponding Organization : University of California, San Francisco
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Variable analysis
independent variables
- Flow rate of bead-containing solution (2000 μl h-1)
- Flow rate of agarose solution (2000 μl h-1)
- Flow rate of 2% ionic Krytox (40,000 μl h-1)
- Formation of monodispersed bead-containing droplets
- Bead material (hydroxylated methacrylic polymer, Toyopearl HW-65S)
- Bead size (20-40 μm)
- Density gradient medium (68% OptiPrep)
- Agarose concentration (2% w/v)
- Agarose type (Ultra-low Gelling Temperature Agarose, Type IX-A)
- Buffer (1× PBS)
- Device height (100 μm)
- Not explicitly mentioned
- Not explicitly mentioned
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