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| Electrophysiological investigation is one of the central tools for studying pore forming proteins. In the past, standard technological approaches have primarily consisted of various alterations of the patch clamp technique of membranes and cells. Here, we introduce an alternative to conventional methods: The Ionovation Compact – a highly flexible bench top system based on the bilayer technique. The bilayer technique is a method to record electrical currents at the single channel level. The bilayer forms a gigaohm resistance between two saline-buffer filled chambers. After incorporation of pore forming proteins (ion channels, solute channels, carriers or pumps), protein mediated currents or membrane potentials can be recorded at high resolution. Stable, low noise, Ag/AgCl electrodes with salt bridges allow recordings in a wide current and salt concentration range. The readily mounted bilayer chamber consists of a 25 µm thick Teflon foil with an aperture of about 50 µm diameter separating two polycarbonate compartments of about 3 ml volume each. The Ionovation Compact system is integrated with the well-known EPC 10 Patch Clamp Amplifier for low noise recording and data acquisition. The Ionovation Compact is controlled by the user-friendly software PATCHMASTER providing full-automated instrument operations, i.e. bilayer production and validation, capacitance control of bilayer integrity (visual control of the bilayer also possible), and perfusion of both membrane sites. A user defined experimental workflow with predefined protocols allows the system to be run in a self-controlled way. |
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