DEs were generated using 3 syringe pumps (PicoPump Elite, Harvard Apparatus) for the inner, oil, and outer carrier fluids. Syringes (1–10 mL; PlastiPak plastic syringes, BD) were connected to the microfluidic device with polyethylene tubing (PE/2, Scientific Commodities). Droplet generation rates were typically 1–10 kHz. For the initial condition (Fig. 2 ), the inner phase for the aqueous droplet core was composed of 1% Tween-20 (Sigma) and FITC-BSA in 1x PBS (Invitrogen). For all measurements, the oil phase was composed of HFE-7500 (Sigma) and 2.2% Ionic PEG-Krytox (FSH 157, Miller-Stephenson) and the outer phase was composed of 1% Tween-20 (Sigma) and 2% Pluronic F68 (Kluplour 188, Sigma) in PBS. Typical flow rates were 400 : 230 : 6500 (O : I : C) μL h−1. Droplet generation was monitored and recorded via a stereoscope (Amscope) and high-speed CMOS camera (ASI 174MM, ZWO). Droplets were stabilized for 4 minutes prior to a set collection time of 6 minutes. At each condition, we acquired a 500-frame video to assess stability and breakoff phenotype.
Hfe 7500
Manufactured by Merck Group
Sourced in United Kingdom, United States
HFE-7500 is a lab equipment product manufactured by Merck Group. It is a nonflammable, low-viscosity, and thermally stable fluorinated fluid designed for use in various laboratory applications. The core function of HFE-7500 is to provide a specialized heat transfer medium for cooling and temperature control purposes in laboratory environments.
Automatically generated - may contain errors
Lab products found in correlation
Fitc bsa, by Thermo Fisher Scientific (3 mentions)
Pluronic f68, by Merck Group (3 mentions)
Tween 20, by Merck Group (3 mentions)
Picopump elite, by Harvard Apparatus (3 mentions)
Plastipak, by BD (2 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions)
Anhydrous dichloromethane, by Merck Group (1 mentions)
Oxalyl chloride, by Merck Group (1 mentions)
Krytox 157fsh, by DuPont (1 mentions)
Polydiallyldimethylammonium chloride, by Merck Group (1 mentions)
Trichloro 1, by Merck Group (1 mentions)
Trichloro 1h, by Merck Group (1 mentions)
5 protocols using hfe 7500
1
Microfluidic Droplet Generation Protocol
2
Synthesis of Biocompatible Fluorinated Surfactant
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The biocompatible fluorinated surfactant used for droplet generation was synthesized as previously described by Chen et al. [27 (link)]. Krytox 157FS (H) (50 g, MW: ~5000 g/mol, Dupont, Londonderry, UK) was dissolved in 50 mL anhydrous HFE-7500 with excess oxalyl chloride (12.5 g, Sigma-Aldrich, Gillingham, UK) and stirred overnight at 85 °C under argon. The solvent was removed through rotary evaporation and high vacuum. The resulting light-yellow product was mixed with Jeffamine XTJ 501 (3.5 g, MW: 900 g/mol, Sigma-Aldrich, UK) and dissolved in a mixture of 50 mL HFE-7500 with 50 mL of anhydrous dichloromethane (50 mL, Sigma-Aldrich, UK) at 65 °C with stirring for 2 days under an argon atmosphere, resulting in a milky white product after rotary evaporation. Insoluble white particles were removed through centrifugation at 8000 rpm for approximately 10 min and dried using a vacuum desiccator for 24 h. Then, the surfactant was used without any further purification.
3
Microfluidic Double Emulsion Generation
4
Microfluidic Double Emulsion Generation
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Double emulsions were generated using 3 syringe pumps (PicoPump Elite, Harvard Apparatus) for the inner, oil, and carrier fluids. The inner phase for the aqueous droplet core was composed of Tween-20 (Sigma) in PBS (Invitrogen), with additional reagents (e.g. FITC–BSA, Invitrogen) as indicated in (Table 1 ). BSA (0.5–2%) can be optionally substituted for Tween-20 (0.1–1%) in the droplet core to no adverse effect. The oil phase was composed of HFE7500 (Sigma) and ionic PEG-Kyrtox teholtze-biocompatible-2008, sukovich-sequence-2017 (FSH, Miller-Stephenson). The carrier phase contained Tween-20 (Sigma) and Pluronic F68 (Kolliphor P 188, Sigma) in PBS. Each phase was loaded into syringes (PlastiPak, BD; Hamilton, Sigma, see ESI† extended methods ), and connected to the device via PE/2 tubing (Scientific Commodities). Typical flow rates were 275 : 75 : 2500 (oil : inner core : outer aqueous sheath) μL h−1. Droplet generation was monitored and recorded via a stereoscope (Amscope) and high-speed CMOS camera (ASI 174MM, ZWO) (Fig. S1† ).
5
Microfluidic Device Fabrication and Surface Modification
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We produce flow focussing microfluidic devices from poly(dimethylsiloxane) (PDMS) (Dow Corning, USA) using soft lithography. 1 The channel walls of single emulsion drop makers are rendered fluorophilic by injecting a HFE-7500-based solution containing 2 vol% of trichloro(1H,1H,2H,2H-perfluorooctyl)silane (Sigma-Aldrich, USA) for 10 min. To modify the surface of double emulsion drop makers, their channel walls are activated with 1M NaOH for 10 min before they are dried with compressed air. The first junction is rendered fluorophilic by treating this part of the channel with a HFE-7500-based solution containing 2 vol% of trichloro(1H,1H,2H,2H-perfluorooctyl)silane (Sigma-Aldrich, USA). The second junction, which is a 3D junction, 2 is rendered hydrophilic using an aqueous solution containing 2 wt.% polydiallyldimethylammonium chloride (Sigma-Aldrich, USA). The solutions are left in the channels for 30 min before channels are dried with compressed air.
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