Automated Patch Clamp Meets High-Throughput Screening: 384 Cells Recorded in Parallel on a Planar Patch Clamp Module

Abstract

We have developed an automated patch clamp module for high-throughput ion channel screening, recording from 384 cells simultaneously. The module is incorporated into a laboratory pipetting robot and uses a 384-channel pipettor head for application of cells and compounds. The module contains 384 amplifier channels for fully parallel recordings using a digital amplifier. Success rates for completed experiments (1- to 4-point concentration-response curves for cells satisfying defined quality control parameters) of greater than 85% have been routinely achieved with, for example, HEK, CHO, and RBL cell lines expressing hNa1.7, hERG, Kir2.1, GABA, or glutamate receptors. Pharmacology experiments are recorded and analyzed using specialized software, and the pharmacology of hNa1.7 and hERG is described. Fast external solution exchange rates of <50 ms are demonstrated using Kir2.1. Short exposure times are achieved by stacking the external solutions inside the pipette of the robot to minimize exposure of the ligand on the receptor. This ensures that ligand-gated ion channels, for example, GABA and glutamate described in this report, can be reproducibly recorded. Stem cell-derived cardiomyocytes have also been used with success rates of 52% for cells that have a seal resistance of >200 MΩ, and recordings of voltage-gated Na and Ca are shown.

Using automated patch clamp electrophysiology platforms in pain‐related ion channel research: insights from industry and academia

“…These drugs (Table ) may produce a clinically useful analgesic or at least provide the research foundations from which such a drug could be successfully developed. The improved recording capabilities that the second generation of APC technology are now providing will be critical to further accelerate the pace of future analgesic drug discovery programmes (Chambers et al , ; Obergrussberger et al , ).…”

“…It is now generally agreed that the most effective and successful of these recording formats was the application of cell suspensions to multiple recording sites on planar recording chips, and became the standard format that subsequent APC platforms would adopt (see Obergrussberger et al , ). In this format, multiple recordings could be achieved simultaneously in a single experimental run.…”

“…For example, the limited fluidics that were used in the first 384‐recording capable APC platform IonWorks, were improved in its later generation IonWorks Barracuda (Gillie et al , ; Kuryshev et al , ). Similarly, Nanion's SyncroPatch 384 PatchEngine (384PE, Obergrussberger et al , ) and Sophion's Qube (Chambers et al , ) also employ micro‐fluidics channels to allow low‐volume, rapid exchange of solutions around the cell recording sites. The second‐generation APC platforms maintain high throughput (384 or 768 simultaneous recordings) whilst achieving the high‐quality GΩ seal recordings, though the 384PE does still require the high‐Ca 2+ ‘seal enhancer’.…”

Human pluripotent stem cell models of cardiac disease: from mechanisms to therapies

“…However, this approach is technically demanding, requiring a skilled operator, and has very low throughput ( Denning et al, 2016 ). Automated patch-clamp platforms can increase throughput, enabling up to 384 cells to be measured simultaneously ( Obergrussberger et al, 2016 ); this method is increasingly used for measuring individual currents in hPSC-CMs, although presently not APs ( Rajamohan et al, 2016 ). A compromise between sensitivity and high-throughput is provided by the multielectrode array (MEA; Box 1 ) ( Asakura et al, 2015 ).…”

Bringing new dimensions to drug discovery screening: impact of cellular stimulation technologies

“…Given that electrophysiology also enables the simultaneous monitoring of cellular responses at a sub-millisecond timescale, it is no wonder that this method is considered the gold standard technology. To make this intricate technology screening friendly, several planar patch-clamp instruments have been developed over the last 15 years [11,12]. The highest throughput (simultaneous acquisition from 384 wells) was achieved in IonWorks Barracuda (Molecular Devices), SyncroPatch 384PE (Nanion), and Qube (Sophion).…”