title: 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 hNaV1.7, hERG, Kir2.1, GABA, or glutamate receptors. Pharmacology experiments are recorded and analyzed using specialized software, and the pharmacology of hNaV1.7 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 Ca2+are shown.
authors.affiliation: Nanion Technologies GmbH, Munich, Germany ali@nanion.de.
authors.forename: Alison
authors.initials: A
authors.lastname: Obergrussberger
authors.affiliation: Nanion Technologies GmbH, Munich, Germany.
authors.forename: Andrea
authors.initials: A
authors.lastname: Brüggemann
authors.affiliation: Nanion Technologies GmbH, Munich, Germany.
authors.forename: Tom A
authors.initials: TA
authors.lastname: Goetze
authors.affiliation: Nanion Technologies GmbH, Munich, Germany.
authors.forename: Markus
authors.initials: M
authors.lastname: Rapedius
authors.affiliation: Nanion Technologies GmbH, Munich, Germany.
authors.forename: Claudia
authors.initials: C
authors.lastname: Haarmann
authors.affiliation: Nanion Technologies GmbH, Munich, Germany.
authors.forename: Ilka
authors.initials: I
authors.lastname: Rinke
authors.affiliation: Nanion Technologies GmbH, Munich, Germany.
authors.forename: Nadine
authors.initials: N
authors.lastname: Becker
authors.affiliation: Nanion Technologies GmbH, Tokyo Laboratory, Medical Innovation Laboratory, Tokyo Women's Medical University and Waseda University Joint Institution for Advanced Biomedical Sciences, Shinjyuku-ku, Tokyo, Japan.
authors.forename: Takayuki
authors.initials: T
authors.lastname: Oka
authors.affiliation: Nanion Technologies GmbH, Tokyo Laboratory, Medical Innovation Laboratory, Tokyo Women's Medical University and Waseda University Joint Institution for Advanced Biomedical Sciences, Shinjyuku-ku, Tokyo, Japan.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Ohtsuki
authors.affiliation: Nanion Technologies GmbH, Munich, Germany.
authors.forename: Timo
authors.initials: T
authors.lastname: Stengel
authors.affiliation: Nanion Technologies GmbH, Munich, Germany.
authors.forename: Marius
authors.initials: M
authors.lastname: Vogel
authors.affiliation: Nanion Technologies GmbH, Munich, Germany.
authors.forename: Juergen
authors.initials: J
authors.lastname: Steindl
authors.affiliation: Nanion Technologies GmbH, Munich, Germany.
authors.forename: Max
authors.initials: M
authors.lastname: Mueller
authors.affiliation: Nanion Technologies GmbH, Munich, Germany.
authors.forename: Johannes
authors.initials: J
authors.lastname: Stiehler
authors.affiliation: Nanion Technologies GmbH, Munich, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: George
authors.affiliation: Nanion Technologies GmbH, Munich, Germany.
authors.forename: Niels
authors.initials: N
authors.lastname: Fertig
mesh_terms: D000818:Animals; D057205:Automation, Laboratory / Q000379:methods*; D002460:Cell Line; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D007473:Ion Channels / Q000032:analysis; D018408:Patch-Clamp Techniques / Q000379:methods*; D011956:Receptors, Cell Surface / Q000032:analysis; D012371:Robotics / Q000379:methods
chemical_list: D007473:Ion Channels; D011956:Receptors, Cell Surface
keywords: automated patch clamp; high-throughput screening; ion channels; ligand-gated ion channels; voltage-gated ion channels