Sonication was used to obtain liposomes of the desired size [26 (link)]. The sonicator (Ultrasonic processor 500 W, Sigma-Aldrich, St. Louis, MO, USA) was set to 40% amplitude and ultrasonic irradiation for 4 cycles (30 s on/60 s off) was applied to liposomal dispersions. An ice bath was used to prevent heating of the dispersions. All sonicated liposomal formulations were stored at 4–8 °C.
Ultrasonic processor 500 w
Manufactured by Merck Group
Sourced in United States
The Ultrasonic processor 500 W is a laboratory instrument used for the disruption and homogenization of samples. It utilizes high-frequency sound waves to agitate and break down the components within a sample, enabling efficient processing and preparation for further analysis or experimentation.
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4 protocols using ultrasonic processor 500 w
1
Liposome Preparation by Sonication
2
Liposome Preparation via Film Hydration Method
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The film hydration method was used to prepare liposomes in aqueous medium [39 (link)]. Briefly, lipids were dissolved in methanol in a round-bottomed flask. To form a thin lipid film, low-pressure rotary evaporation was performed on a Büchi Rotavapor R-124 with vacuum pump V-700 (Büchi Labortechnik, Flawil, Switzerland). The film was then resuspended in 8 mL of isotonic phosphate buffer (pH 7.4, 290 mOsm) by hand-shaking and 10 min bath sonication (Bransonic® ultrasonic 5510, Vlierberg, Holland). A concentration of 10 mg/mL of neutral SPC S100 (with over 94% of pure phosphatidylcholine from soybean) was used for all formulations. Fluorescent dyes (T and C) were separately incorporated in the initial lipid mixture at the concentrations of 0.015, 0.030, 0.060 and 0.120 mg/mL, respectively.
The effective size reduction of the multilamellar dispersions was achieved by combining 2 min sonication (Ultrasonic processor 500 W, Sigma-Aldrich, MO, USA) and a stepwise hand extrusion through Nucleopore® polycarbonate membranes (with sieving sizes of 400 and 200 nm, respectively). Overnight stabilization was allowed in between the steps and prior to the characterization. All liposomal suspension were then stored in the fridge (4 °C) and at room temperature (25 °C), respectively.
The effective size reduction of the multilamellar dispersions was achieved by combining 2 min sonication (Ultrasonic processor 500 W, Sigma-Aldrich, MO, USA) and a stepwise hand extrusion through Nucleopore® polycarbonate membranes (with sieving sizes of 400 and 200 nm, respectively). Overnight stabilization was allowed in between the steps and prior to the characterization. All liposomal suspension were then stored in the fridge (4 °C) and at room temperature (25 °C), respectively.
3
Liposome Size Reduction by Probe Sonication
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The vesicle size of the liposomes was reduced by probe sonication [28 (link)]. The liposomal suspension was placed on ice and the needle probe tip inserted approximately 5–7 mm into the liposomal suspension. The sonicator (Ultrasonic processor 500 W, Sigma–Aldrich, St. Louis, MO, USA) was set to 40% amplitude and the samples sonicated 3 times for 1 min, with 1 min resting periods. The samples were stored in the refrigerator (4–6 °C) for at least 6 h prior to further experiments.
4
Sonication-Induced Liposome Preparation
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Liposomes (5 mL; 49.5 mg/mL or 54.0 mg/mL, respectively) from each formulation was transferred to 10 mL beaker and placed on ice bath. The sonicator (Ultrasonic processor 500 W, Sigma-Aldrich, St. Louis, MO, USA) was set to 40% amplitude and the liposomes were exposed to ultrasonic irradiation for 1, 2 or 2 × 2 min, respectively, to obtain large and small
liposomes of approximately 160 nm and 50 nm in diameter, respectively. The sonicated liposomes were stored in the refrigerator for at least 6 h prior to further use. The free RBITC (Mw 536.08) was separated from liposomally entrapped RBITC by using Nanosep 100K
(MWCO 100 KDa) modified polyether sulphone ultrafiltration devices (Pall life Sc, NY, USA).
The supernatant, liposomal suspension free of unentrapped RBITC, was used for further characterization. The particle size distributions of liposomes were determined by photon correlation spectroscopy (PCS) (Submicron particle sizer model 370, Nicomp, Santa Barbara, CA, USA) and zeta potential measurements were performed on a Malvern Zetasizer Nano Z (Malvern, Oxford, UK) as described according to Jøraholmen et al. (Jøraholmen et al., 2015) .
The liposomes were stored at 4 °C and used in experiments within 7 days after preparation.
liposomes of approximately 160 nm and 50 nm in diameter, respectively. The sonicated liposomes were stored in the refrigerator for at least 6 h prior to further use. The free RBITC (Mw 536.08) was separated from liposomally entrapped RBITC by using Nanosep 100K
(MWCO 100 KDa) modified polyether sulphone ultrafiltration devices (Pall life Sc, NY, USA).
The supernatant, liposomal suspension free of unentrapped RBITC, was used for further characterization. The particle size distributions of liposomes were determined by photon correlation spectroscopy (PCS) (Submicron particle sizer model 370, Nicomp, Santa Barbara, CA, USA) and zeta potential measurements were performed on a Malvern Zetasizer Nano Z (Malvern, Oxford, UK) as described according to Jøraholmen et al. (Jøraholmen et al., 2015) .
The liposomes were stored at 4 °C and used in experiments within 7 days after preparation.
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