Validation of an Automated Electrophysiology System that Measures Ligand- or Voltage-Gated Ion Channels Simultaneously in 384 Wells

Abstract

We report in this study the validation of an automated patch clamp system that enables a novel way of analyzing ligand-gated ion channel (LGIC) pharmacology, based on high uniformity of currents at 384 parallel recording wells. The system combines a 384-well pipettor with 384 amplifiers and digitizers for parallel recording in all wells at once, no multiplexing is performed. The system measures cell membrane currents using the perforated patch clamp technique on a polyimide substrate. Currents are measured using either a single hole (SH) in each recording well or an array of 64 holes in each well (Population Patch Clamp or PPC). In this study we validated the performance of IonWorks Barracuda™ system with multiple ion channels targets. Data presented here include measurement of solution exchange rate, LGIC recordings of nicotinic acetylcholine receptors α1 and α7, acid sensing ion channels, and GABA chloride channels; as well as VGIC recordings of NaV, KV and hERG channels. Pharmacological blockade of ion channel activity is also presented to validate the use of this automated, high-throughput system for screening ion channel targets in a drug discovery setting.