DSPC, DSPG, and cholesterol
were added into a 50 mL round-bottom flask at a 1:1:1 mol/mol/mol
ratio. Dried thin films were created by heating the round-bottom at
50 °C, spinning, and pumping the pressure down to <25 mTorr
for at least 1 h using a rotary evaporator system (Buchi). For NPs
used in confocal microscopy, 0.4 mol % DSPE-Cy5 lipid was added to
replace DSPC, resulting in a 32.9:33.3:33.4:0.4 DSPC/DSPG/Chol/DSPE-Cy5
molar ratio.
After the evaporation of all organic solvents,
the round-bottomed flask was partially submerged in a sonicator bath
(Branson 1800 Ultrasonic Bath) filled with 65 °C water, and the
thin film was resuspended at 1 mg/mL using fresh 65 °C milli-Q
water while sonicating. The resulting colloidal suspension was extruded
at 65 °C by using an Averin LipsosFast LF-50 liposome extruder.
Liposomes were first passed three times through a 200 nm filter, then
three times through a 100 nm filter, and finally at least once through
a 50 nm filter.
were added into a 50 mL round-bottom flask at a 1:1:1 mol/mol/mol
ratio. Dried thin films were created by heating the round-bottom at
50 °C, spinning, and pumping the pressure down to <25 mTorr
for at least 1 h using a rotary evaporator system (Buchi). For NPs
used in confocal microscopy, 0.4 mol % DSPE-Cy5 lipid was added to
replace DSPC, resulting in a 32.9:33.3:33.4:0.4 DSPC/DSPG/Chol/DSPE-Cy5
molar ratio.
After the evaporation of all organic solvents,
the round-bottomed flask was partially submerged in a sonicator bath
(Branson 1800 Ultrasonic Bath) filled with 65 °C water, and the
thin film was resuspended at 1 mg/mL using fresh 65 °C milli-Q
water while sonicating. The resulting colloidal suspension was extruded
at 65 °C by using an Averin LipsosFast LF-50 liposome extruder.
Liposomes were first passed three times through a 200 nm filter, then
three times through a 100 nm filter, and finally at least once through
a 50 nm filter.