Efficient use of membrane protein nanopores in ionic single-molecule sensing requires technology for the reliable formation of suspended molecular membranes densely arrayed in formats that allow high-resolution electrical recording. Here, automated formation of bimolecular lipid layers is shown using a simple process where a poly(tetrafluoroethylene)-coated magnetic bar is remotely actuated to perform a turning motion, thereby spreading phospholipid in organic solvent on a nonpolar surface containing a <1 mm(2) 4 × 4 array of apertures with embedded microelectrodes (microelectrode cavity array). Parallel and high-resolution single-molecule detection by single nanopores is demonstrated on the resulting bilayer arrays, which are shown to form by a classical but very rapid self-assembly process. The technique provides a robust and scalable solution for the problem of reliable, automated formation of multiple independent lipid bilayers in a dense microarray format, while preserving the favorable electrical properties of the microelectrode cavity array.

During bursts in KO, the rate was comparable to WT (4.150.3Hz). WT SANs (n=5) showed a 70.556.1% increase in rate after exposure to the b-adrenergic agonist ISO (10mM), while KO showed no increase in average rate. However, when considering only rate during burst activity in KO, 6 out of 10 KO SANs responded significantly to ISO 10mM (52516% increase). The specific I f inhibitor ivabradine (IVA, 9mM) reduced the spontaneous pacemaker rate in both WT (n=6) and KO (n=8) SANs (4459% and 58.956.8% decrease, respectively), and even during the bursts in the KO (36517.9% decrease). Thus, NCX1 KO SANs can generate bursts of pacemaker activity similar to WT. These bursts are responsive to both ISO and IVA, consistent with I f-mediated pacemaker activity despite the absence of NCX.