3 axis water hydraulic micromanipulator

A schematic of the experimental set-up for forming supported bilayer patches is shown in Fig. 1. A laser scanning confocal microscope (Leica TCS SP8, Leica Microsystems GmbH, Wetzlar, Germany) was used. The supported lipid bilayers were formed in situ by transforming SUVs on a glass substrate (WillCo Wells B.V. Amsterdam, NL), using an open-space microfluidic multichannel pipette 17 (Fluicell AB, Sweden). For deposition of the lipids, the microfluidic pipette was positioned using a 3-axis water hydraulic micromanipulator (Narishige, Japan) 10-20 mm above the surface and the recirculation of SUVs (0.1 mg ml À1 ) was initiated (Fig. 1a). This leads to the adhesion of SUVs onto the solid surface, rupturing and eventual merging of the individual ruptured lipid patches into a circular homogeneous planar bilayer 17 (Fig. 1a). Approximately 2 minutes after forming the lipid patch, this time SMA copolymer was applied to the newly formed bilayer via the pipette which in some instances lead to pore formation (Fig. 1b). All experiments were performed at constant room temperature of 18 1C.

A schematic of the experimental set-up for forming supported bilayer patches is shown in Fig. 1. A laser scanning confocal microscope (Leica TCS SP8, Leica Microsystems GmbH, Wetzlar, Germany) was used. The supported lipid bilayers were formed in situ by transforming SUVs on a glass substrate (WillCo Wells B.V. Amsterdam, NL), using an open-space microfluidic multichannel pipette 17 (Fluicell AB, Sweden). For deposition of the lipids, the microfluidic pipette was positioned using a 3-axis water hydraulic micromanipulator (Narishige, Japan) 10-20 mm above the surface and the recirculation of SUVs (0.1 mg ml À1 ) was initiated (Fig. 1a). This leads to the adhesion of SUVs onto the solid surface, rupturing and eventual merging of the individual ruptured lipid patches into a circular homogeneous planar bilayer 17 (Fig. 1a). Approximately 2 minutes after forming the lipid patch, this time SMA copolymer was applied to the newly formed bilayer via the pipette which in some instances lead to pore formation (Fig. 1b). All experiments were performed at constant room temperature of 18 1C.