Production of Nanoemulsions with Microfluidization

All nanoemulsions were produced following earlier published protocols^{18 (link),33 (link),36 (link),38 (link)}. Briefly, a micelle solution of blended non-ionic surfactants (2% Pluronic P-105, 3% Pluronic P-123 w/v) was made by following previously reported protocol with minor modifications^{55 (link)}. To prepare nanoemulsions, micelle solution was added to the mixture of PFC and HC oils, or single (PFC or HC) oil. Then, coarse pre-emulsions were produced with an analog vortex mixer (VWR, Radnor, PA, USA) on high for 30 s. Coarse emulsions were sonicated on ice for 30 s at 29% amplitude (equivalent of 3480 W s) with Model 450 Digital Sonifier (BRANSON Ultrasonics Corporation, Danbury, CT, USA). Microfluidization was performed on all coarse emulsions by Microfluidizer M110S (Microfluidics Corp., Westwood, MA, USA) at 15,000 psi liquid pressure for the specified number of pulses (5 pulses to 1 pass) over an ice-cold interaction chamber. Emulsions were packaged in glass vials and stored at 4 °C.