| Autor: |
Obergrussberger, Alison, Brüggemann, Andrea, Goetze, Tom A., Rapedius, Markus, Haarmann, Claudia, Rinke, Ilka, Becker, Nadine, Oka, Takayuki, Ohtsuki, Atsushi, Stengel, Timo, Vogel, Marius, Steindl, Juergen, Mueller, Max, Stiehler, Johannes, George, Michael, Fertig, Niels |
| Zdroj: |
Journal of Laboratory Automation; Dec2016, Vol. 21 Issue 6, p779-793, 15p |
| Abstrakt: |
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. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Supplemental Index |
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