Scanning tunneling microscopy is based on the flow of an electrical current and thus cannot be used to directly image insulating material. It has been found, however, that a very thin film of water (about one monolayer) adsorbed to a surface exhibits a surprisingly high conductivity that is sufficient to allow scanning tunneling microscope imaging at currents below 1 picoampere. Hydrophilic insulators, such as glass and mica, can thus be imaged in humid air. The same is true for biological specimens deposited on such surfaces, as demonstrated by the scanning tunneling microscope imaging of plasmid DNA on mica.
Lateral electric conductivity of mica-supported lipid monolayers and of the corresponding lipid bilayers has been studied by means of scanning tunneling microscopy (STM). The surface of freshly cleaved mica itself was found to be conductive when exposed to humid air. Lipid monolayers were transferred onto such a surface by means of the Langmuir-Blodgett technique, which makes the mica surface hydrophobic and suppresses the electric current along the surface in the experimentally accessible humidity (5-80%) and applied voltage (0-10 V) range. This is true for dipalmitoylphosphatidylethanolamine (DPPE) as well as dipalmitoylphosphatidylcholine (DPPC) monolayers. Repeated deposition of DPPC layers by means of the Langmuir-Blodgett LB technique does not lead to the formation of a stable surface-supported bilayer because of the high hydrophilicity of the phosphatidylcholine headgroups that causes DPPC/DPPC bilayers to peel off the supporting surface during the sample preparation. In contrast to this, a DPPE or a DPPC monolayer on top of a DPPE monolayer gives rise to a rather stable mica-supported bilayer that can be studied by STM. Electric currents between 10 and 100 fA, depending on the ambient humidity, flow along the DPPE bilayer surface, in the humidity range between 35 and 60%. The DPPC surface, which is more hydrophilic, is up to 100 times more conductive under comparable conditions. Anomalous high lateral conductivity thus depends on, and probably proceeds via, the surface-adsorbed water layers. The prominence of ambient humidity and surface hydrophilicity on the measured lateral currents suggests this. The combination of our STM data and previously published water adsorption isotherms as a function of the relative humidity indicate that one layer or less of adsorbed water suffices for mediating the measurable lateral currents. The fact that similar observations are also made for other hydrophilic substrates supports the conclusion that lateral conductivity via surface-adsorbed water is a rather general phenomenon.
A simple method for rendering atomic force microscope tips and cantilevers hydrophilic or hydrophobic through glow discharge in an appropriate gas atmosphere is introduced. Force curves at different humidities of these modified cantilevers were taken on freshly cleaved mica (hydrophilic surface) and on a monolayer of dipalmitoylphosphatidylethanolamine transferred onto mica (hydrophobic surface) to characterize the behavior of the cantilevers on hydrophilic and hydrophobic surfaces. Furthermore, Langmuir-Blodgett bilayers, with a dipalmitoylphosphatidylethanolamine bottom layer and a dipalmitoylphosphatidylcholine top layer, were imaged in the constant force mode in a multimode atomic force microscope in air under controlled humidity conditions. The friction and elasticity signal were recorded parallel to the topography. By varying the force exerted by the tip on the sample, different layers of the Langmuir-Blodgett system could be removed or flattened. Removal exposed underlying layers that exhibited a different friction and elasticity behavior. Furthermore, force scans with tips rendered hydrophobic were taken on the different layers of the sample to characterize the hydrophilic/hydrophobic nature of the layers. Only by combining the results obtained by the different methods can the structure of the lipid layer systems be identified.
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