Optogenetics and Optical Tools in Automated Patch Clamping

Methods Mol Biol. 2021:2188:311-330. doi: 10.1007/978-1-0716-0818-0_16.

Abstract

Automated patch clamping (APC) has been used for almost two decades to increase the throughput of electrophysiological measurements, especially in preclinical safety screening of drug compounds. Typically, cells are suctioned onto holes in planar surfaces and a stronger subsequent suction allows access to a whole cell configuration for electrical measurement of ion channel activity. The development of optogenetic tools over a wide range of wavelengths (UV to IR) provides powerful tools for improving spatiotemporal control of in vivo and in vitro experiments and is emerging as a powerful means of investigating cell networks (neuronal), single cell transduction, and subcellular pathways.Combining APC and optogenetic tools paves the way for improved investigation and control of cell kinetics and provides the opportunity for collecting robust data for new and exciting applications and therapeutic areas. Here, we present an APC optogenetics capability on the Qube Opto 384 system including experiments on light activated ion channels and photoactivated ligands.

Keywords: APC; Automated patch clamp; Caged compounds; ChR2; Channelrhodopsin; Fast ligand gated channels; HCN phosphorylation; Optogenetics; Qube Opto 384; Rubi-GABA; Spatiotemporal kinetics; bPAC; cAMP.

MeSH terms

  • Cell Culture Techniques / methods
  • Channelrhodopsins / genetics
  • Channelrhodopsins / metabolism
  • HEK293 Cells
  • Humans
  • Ion Channels / genetics
  • Ion Channels / metabolism
  • Light
  • Optogenetics / methods*
  • Patch-Clamp Techniques / methods*

Substances

  • Channelrhodopsins
  • Ion Channels