Tension-stabilized pores in giant vesicles: determination of pore size and pore line tension

Abstract

We present the first observations of giant, long-existing, stabilized pores in vesicle membranes. Using a new experimental technique for studying the electropermeabilization of lipid membranes, giant liposomes (from 25 to 56 μm in diameter) were subjected to single, square, electric pulses (duration 150 μs and electric field strength from 63 to 126 kV/m). The liposomes were held by a micropipet and small membrane tensions were created by controlling the pipet suction pressure. The liposomes were loaded with media having different refractive index from the outside solution, and, under these conditions, the formation of pores in the pressurized liposome could be visualized by the jet of inside solution that flowed out from the membrane pore. By adjusting the membrane tension, pores were kept open, and pore lifetimes could be varied from tenths of a second to several seconds. The pore size was determined from the volumetric flow in the pore region and the measured pressure differences across the bilayer. It was clear from the experiments that only one pore remained opened after the pulse. The estimated pore radii were on the order of one micrometer. The pores were in a quasi-stationary state and when they closed they did so spontaneously in a quick process (in milliseconds). The isotropic membrane tension was determined for the same measurements and from determinations of both pore size and dynamic membrane tension the pore line tension was found. The line tension of the pore region was determined for two lipid compositions, stearoyloleoylphosphatidylcholine and stearoyloleoylphosphatidylcholine with 50 mol% cholesterol, and the obtained values for single bilayers were (0.92 ± 0.07)·10 −11 N and (3.05 ± 0.12)·10 −11 N, respectively.