A Novel High-Throughput Combined Voltage-Clamp/Current-Clamp Analysis of Single Primary Neurons




Mohammad-Reza Ghovanloo, Sidharth Tyagi, Peng Zhao, Emre Kiziltug, Mark Estacion, Sulayman D. Dib-Hajj, and Stephen G. Waxman



The patch-clamp technique is the gold-standard methodology for biophysical investigations of channels
and receptors studied by voltage-clamp, and for analysis of excitability of cells, such as neurons studied
via current-clamp. However, the throughput of manual patch-clamp is slow, and high-throughput robotic
patch-clamp, while helpful for assessments such as drug screening, has been primarily used to study
channels and receptors expressed in heterologous expression systems. In this study, we introduce a
novel approach to automated high throughput patch-clamping that substantially enhances high
throughput analysis of excitable cells at the channel and cellular level. As a proof-of-concept, we apply
this approach to investigate the detailed biophysical properties of voltage-gated sodium (Nav) channels
in dorsal root ganglion (DRG) neurons, which are among the most diverse and complex neuronal cells.
Our approach enables high throughput, unbiased, fast, simultaneous, and head-to-head
electrophysiological recordings from a wide range of primary neurons. Furthermore, our approach
eliminates the need for culturing of cells on coverslips and provides the ability to perform both voltage and
current-clamp recordings on the same neuron. This approach can be used for many applications,
including both physiological and pharmacological analyses of primary DRG neurons.

Key words: Current clamp

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