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2021 - The suitability of high throughput automated patch clamp for physiological applications

icon sp96  SyncroPatch 384 Publication in The Journal of Physiology (2021)

Authors:
Obergrussberger A., Rinke-Weiß I., Goetze T.A., Rapedius M., Brinkwirth N., Becker N., Rotordam M.G., Hutchison L., Madau P, Pau D, Dalrymple D., Braun N., Friis S., Pless S.A., Fertig N.

Journal:

The Journal of Physiology (2021) doi:10.1113/JP282107


Abstract: 

Although automated patch clamp (APC) devices have been around for many years and have become an integral part of many aspects of drug discovery, high throughput instruments with GΩ seal data quality are relatively new. Experiments where a large number of compounds are screened against ion channels are ideally suited to high throughput APC, particularly when the amount of compound available is low. Here we evaluate different APC approaches using a variety of ion channels and screening settings. We have performed a screen of 1,920 compounds on GluN1/GluN2A NMDA receptors for negative allosteric modulation using both the SyncroPatch 384 and FLIPRTM. Additionally, we tested the effect of 36 arthropod venoms on NaV1.9 using a single 384-well plate on the SyncroPatch 384. As an example for mutant screening, a range of acid-sensing ion channel variants were tested and the success rate increased through FACS prior to APC experiments. GΩ seal data quality makes the 384- format accessible to recording of primary and stem cell-derived cells on the SyncroPatch 384. We show recordings in voltage and current clamp modes of stem cell-derived cardiomyocytes. In addition, the option of intracellular solution exchange enabled investigations into the effects of intracellular Ca2+ and cAMP on TRPC5 and HCN2 currents, respectively. Together, this highlights the broad applicability and versatility of APC platforms and also outlines some limitations of the approach.


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