A High Throughput Microfluidic Approach Enables Fast Exchange Of Solutions And Ligand Gated Ion Channel Recording From Cell Ensemble

Abstract:

Electrophysiology is the preferred technique for characteriz- ing ion channel function and kinetics. It is the most function- ally pertinent assay for screening in terms of information content. High throughput pharmaceutical screens often use a population patch approach, which eliminates cell-to cell vari- ability of single cell recordings. However, currently available population patch platforms have key shortcomings such as a) the inability for fast exchange of solutions, b) the inability to apply multiple compounds to the same ensemble of cells, and c) the inability to record fast desensitizing channels.

Here we present novel data showing that by using a microflu- idic network design along with population patch recording we are able to overcome these obstacles. We validated our system using cells expressing voltage-gated channels in en- sembles of up to 30 cells under voltage clamp. Moreover, these results showed that there is fast compound application (<100ms). The time course of compound application was con- firmed using fluorescent indicators and biological reporters, such as GABA-A expressing cells. These data also validated our ability to record from fast desensitizing ligand gated ion channels without appreciable desensitization. We compared the time course of solution exchange with and without a pro- tective layer technique and additionally characterized appli- cation of multiple compounds to the same ensemble of cells. In conclusion, the novel microfluidic approach allows for the fast exchange of compounds and facilitates the recording of fast activating voltage and ligand-gated channels.

 

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