IonFlux: A Microfluidic Approach To Ion Channel Ensemble Recording For Ligand-Gated And Voltage-Gated Ion Channels

 

Jeff Jensen, C. Ian Spencer, Nianzhen Li, Juliette Johnson, Qin Chen, and Cristian Ionescu-Zanetti
Fluxion Biosciences, South San Francisco, CA

Abstract:

Automated patch clamp (APC) addresses a need for high throughput screening of chemical entities altering ion channel function. Systems that can produce pharmacologically relevant data increasingly in academic laboratories. Here we present data obtained with a novel APC platform rates no internal robotic liquid handling, and features continuous recording from cell ensembles during rapid solution switches with a bench-top footprint resembling a conventional plate reader.

True whole cell voltage clamp was applied to linear arrays of twenty cells in parallel, utilizing fully-featured 16 or 64 channel voltage-clamp amplifiers uder computer control. Laminar flow of solutions in a microfluidic network delivered cells in suspension to the recording sites and en- abled fast exchange of bathing solutions via an electro-pneumatic interface, on standard well plate formats. Electrophysiological characterization is presented for hERG and KV potassiu chan- nels, NaV sodium channels, and GABA receptor channels. Our results show both voltage- dependence of theses currents and their modulation by pharmacological agents. The recordings also demonstrate the potential for microfluidic-enabled, fast perfusion and washout of candidate drugs. Incoporation of multiple experiments per well-plate enables a large number of com- pounds to be profiled in parallel, especially if multiple distict compounds are applied to each cell ensamble. Data on recording success raes, throughput, and assay reproducibility show that high throughput experiments can be performed with enhanced reliability

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