PFC-NEs for in vivo and ex vivo fluorescent tracking of monocyte derived macrophages used in this study were prepared by adapting earlier published methods [27 (link), 28 (link)]. PFC-NEs were formulated to incorporate two fluorescent dyes in the hydrocarbon phase. Perfluoro-15-crown-5 ether (14.2% w/v) was blended by vortexing with Miglyol 812 N (7.6% w/v) which incorporated two dyes: 1,1′-Dioctadecyl-3,3,3′,3′-Tetramethylindocarbocyanine Perchlorate [‘DiI’; DiIC18(3)], at 10 μM and (‘DiR’; 1,1′-Dioctadecyl-3,3,3′,3′-Tetramethylindotricarbocyanine Iodide, DiIC18(7)), at 10 or 25 μM final concentration, and with fluorescence emission maxima at 570 nm and 780 nm respectively. The oils were mixed with non-ionic surfactant solution prepared in 1X phosphate buffered saline (PBS) and processed on Microfluidizer M110S (Microfluidics, Westwood, MA 02090, USA) at 18,000 psi liquid pressure on 25 mL scale as reported earlier [27 (link)]. All nanoemulsions are sterilized by filtration through a 0.22 μm filter and stored in sterilized amber vials before use.
Microfluidizer m110s
Manufactured by Microfluidics
Sourced in United States
The Microfluidizer M110S is a high-pressure homogenizer designed for processing fluid samples. It utilizes a high-pressure interaction chamber to create a uniform and controlled emulsion or dispersion of materials within a liquid.
Automatically generated - may contain errors
6 protocols using microfluidizer m110s
1
Fluorescent Tracking of Monocyte-Derived Macrophages
2
Citrus Fiber-Based Emulsion Formulation
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The citrus fiber powder was first suspended in deionized water and thoroughly mixed using a LM5-A Silverson laboratory mixer (Silverson, USA) with a 1 mm screen hole at 3500 rpm for 5 minutes and afterwards passed once through a high-pressure homogenizer (Microfluidizer M 110S (MF), Microfluidics Corp., USA) with a z-shape geometry (diameter 87 μm) operating at a pressure of 1200 bar. Then soybean oil was added, and the mixture was once again mixed using the Silverson at 3500 rpm for 5 minutes and then microfluidized at 1200 bar. Samples were prepared with 0.5; 1.0; 1.5; 2.0 wt % plant cell wall dispersion in the water phase, hence concentrations in CMF are respectively 0.3; 0.6; 0.9; 1.2 wt%. Oil concentrations in the emulsions are 15; 30; 40; 50; 60 wt % soybean oil in total, all concentrations reported are according to these. All samples were stored in closed containers at 5 °C after preparation.
3
Nanoemulsion Preparation via Microfluidization
4
Microfluidized Kolliphor EL/Pluronic P105 Emulsions
5
Production of Nanoemulsions with Microfluidization
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All nanoemulsions were produced following earlier published protocols18 (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 modifications55 (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.
6
High-Pressure Nanoemulsion Preparation
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Microfluidizer M110S and Microfluidizer M-110EH-30 from Microfluidics Corporation, Westwood, Massachusetts, were used for all nanoemulsion preparations.
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