Screening Fluorescent Voltage Indicators with Spontaneously Spiking HEK Cells |
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| Authors: | Jeehae Park Christopher A. Werley Veena Venkatachalam Joel M. Kralj Sulayman D. Dib-Hajj Stephen G. Waxman Adam E. Cohen |
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| Affiliation: | 1. Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, United States of America.; 2. Department of Neurology, Yale School of Medicine, and Neurorehabilitation Research Center, Veterans Affairs Hospital, West Haven, Connecticut, United States of America.; 3. Department of Physics, Harvard University, Cambridge, Massachusetts, United States of America.; Brigham & Women''s Hospital - Harvard Medical School, United States of America, |
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| 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. |
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