Water in Fluorocarbon Oil Emulsions

Water in fluorocarbon oil emulsions (W/FO) are widely used in R&D environments primarily to achieve compartmentalization of biological samples within droplets and prevent egress of the material to the surrounding media such as Drop-Seq and RNA-Seq.  Of particular importance is the ability to produce high-quality, monodisperse droplets and the ability to do so reproducibly and at a high production rate.

The combination of Fluigent pressure pump units and Raydrop microfluidic devices developed and manufactured by Secoya enable smooth fluid delivery, precision flowrate control, automation, and reproducibility necessary to generate high-quality water in fluorocarbon oil emulsions.

Developed and manufactured by Secoya

Materials and methods

Reagents

Droplet Phase: Water (Mili Q)

Continuous phase: dSurf

ReagentSupplierCatalogue numberCAS Number
WaterUltrapure 18.2 MΩ – cm7732-18-5
dSURFFluigentDR-RE-SU-12

Microfluidic Setup

The microfluidic setup was composed of:

Protocols steps

Water-in-fluorocarbon-Scheme of the fluidic setup
Figure 1: Scheme of the fluidic setup
Pictures of the Fluigent equipment
Figure 2: Pictures of the Fluigent equipment

Results

Continuous phase flowrate
(μl/min)
Droplet phase flowrate
(μl/min)
Droplet diameter
(μm)
Production rate
(Hz)
100563636
10010671058
10015711334
50568506
501073818
5015761087
25572426
251075754
251581898
151585777
Water-in-Fluorocarbon-droplet phase diagram
Figure 3: Droplet phase diagram
WATER-in-Fluorocarbon-water droplets in dSURF
Figure 4: Images of water droplets in dSURF generated using Fluigent equipment and Raydrop microfluidic device

Conclusion

Fluigent pressure-based flow controller units and Raydrop microfluidic device were successfully used to generate high-quality, monodisperse droplets of water in fluorocarbon oil. The droplet size was controlled in the range of 63 – 85 μm by adjusting the continuous and dispersed phase flowrates. Peak stable droplet production rate was recorded for 71 μm droplets at 1334 Hz. The production techniques developed here can use a wide range of compartmentalization applications such as Drop-Seq and RNA-Seq

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