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Screening fluorescent voltage indicators with spontaneously spiking HEK cells

Screening fluorescent voltage indicators with spontaneously spiking HEK cells

Plos One 8(12): E85221

Development of improved fluorescent voltage indicators is a key challenge in neuroscience, but progress has been hampered by the low throughput of patch-clamp characterization. We introduce a line of non-fluorescent HEK cells that stably express NaV 1.3 and KIR 2.1 and generate spontaneous electrical action potentials. These cells enable rapid, electrode-free screening of speed and sensitivity of voltage sensitive dyes or fluorescent proteins on a standard fluorescence microscope. We screened a small library of mutants of archaerhodopsin 3 (Arch) in spiking HEK cells and identified two mutants with greater voltage-sensitivity than found in previously published Arch voltage indicators.

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Accession: 055670663

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PMID: 24391999

DOI: 10.1371/journal.pone.0085221

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