Patch-clamping of primary cardiac cells with micro-openings in polyimide films

Patch-clamping is a powerful method for investigating the function and regulation of ionic channels. Currently, great efforts are being made to automate this method. As a step towards this goal, the feasibility of patch-clamping primary cells with a microscopic opening in a planar substrate was tested. Using standard microfabrication and ion beam technology, small-diameter openings (2 and 4 mum) were formed in polyimide films (thickness 6.5 mum). Single cells (sheep Purkinje heart cells, Chinese hamster ovary cells) in a suspension were positioned on top of the opening and sucked towards the opening to improve adhesion of the cell to the planar substrate, hence increasing the seal resistance. Voltage/current measurements yielded a median seal resistance of 1.3MOmega with 4 mum openings (n=24) and 26.0 MOmega with 2mum openings (n=75), respectively. With 2mum openings, successful loose-patch recordings of TTX-sensitive inward currents and action potentials in sheep Purkinje heart cells (n = 18) were made. In rare cases, gigaseals (n = 4) were also measured, and a whole-cell configuration (n=1) could be established. It was concluded that the simple planar patch approach is suitable for automated loose-patch recordings from cells in suspension but will hardly be suitable for high-throughput whole-cell patch-clamping with high-resistance seals.