Frozen-hydrated liposome samples for cryo-TEM with varied HIP content (0-600 µg/ml) were prepared at a concentration of 10 mg/ml in 100 mM phosphate buffer (pH 7.4) and 4 μl of each sample were deposited on glow-discharged C-Flat™ Holey Carbon CF-224C 400 Mesh TEM grids (Electron Microscopy Sciences) in the chamber of the Leica GP2 plunger. The chamber was set to 20 °C and 40% humidity. After 10 s incubation, the grids were blotted for 2-3 s (with 2 mm push and without additional drain time) and plunge-frozen in liquid ethane maintained at -183 °C by liquid nitrogen.