The suitability of high throughput automated patch clamp for physiological applications

Area of Science:

• Pharmacology
• Neuroscience
• Biophysics

Background:

• Automated patch clamp (APC) is crucial for drug discovery, with recent high-throughput instruments achieving gigaohm seal quality.
• High-throughput APC is ideal for screening numerous compounds against ion channels, especially with limited compound availability.

Purpose of the Study:

• Evaluate diverse APC approaches for ion channel screening across various settings.
• Demonstrate the utility of high-throughput APC for compound screening, venom analysis, and genetic variant studies.

Main Methods:

• Screened 1920 compounds on GluN1/GluN2A NMDA receptors using SyncroPatch 384 and FLIPR.
• Tested 36 arthropod venoms on Na_V 1.9 using SyncroPatch 384.
• Utilized fluorescence-activated cell sorting (FACS) for mutant screening of acid-sensing ion channels prior to APC.
• Recorded from stem cell-derived cardiomyocytes in voltage and current clamp modes.
• Investigated intracellular ion and second messenger effects using intracellular solution exchange.

Main Results:

• SyncroPatch 384 demonstrated lower false positive rates than FLIPR in high-throughput NMDA receptor screening.
• Successful screening of genetic variants and mutations on a single 384-well plate.
• Achieved successful recordings from primary and stem cell-derived cells, including action potentials.
• Demonstrated investigation of intracellular Ca²⁺ and cAMP effects on TRPC5 and HCN2 currents.

Conclusions:

• High-throughput APC platforms are broadly applicable and versatile for ion channel research.
• APC facilitates efficient screening, genetic variant analysis, and intracellular studies with high data quality.
• APC platforms are suitable for recordings from primary and stem cell-derived cells, expanding their utility in drug discovery and research.