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Screening fluorescent voltage indicators with spontaneously spiking HEK cells.

ABSTRACT: 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.

SUBMITTER: Park J

PROVIDER: S-EPMC3877367 | biostudies-literature | 2013

REPOSITORIES: biostudies-literature

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Screening fluorescent voltage indicators with spontaneously spiking HEK cells.

Park Jeehae J Werley Christopher A CA Venkatachalam Veena V Kralj Joel M JM Dib-Hajj Sulayman D SD Waxman Stephen G SG Cohen Adam E AE

PloS one 20131231 12

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 ...[more]

PMID: 24391999

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