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Hochbaum, Daniel R; Zhao, Yongxin; Farhi, Samouil L; Klapoetke, Nathan; Werley, Christopher A; Kapoor, Vikrant; Zou, Peng; Kralj, Joel M; Maclaurin, Dougal; Smedemark-Margulies, Niklas; Saulnier, Jessica L; Boulting, Gabriella L; Straub, Christoph; Cho, Yong Ku; Melkonian, Michael; Wong, Gane Ka-Shu; Harrison, D Jed; Murthy, Venkatesh N; Sabatini, Bernardo L; Boyden, Edward S; Campbell, Robert E; Cohen, Adam E
Nature methods, 08/2014, Letnik: 11, Številka: 8Journal Article
All-optical electrophysiology-spatially resolved simultaneous optical perturbation and measurement of membrane voltage-would open new vistas in neuroscience research. We evolved two archaerhodopsin-based voltage indicators, QuasAr1 and QuasAr2, which show improved brightness and voltage sensitivity, have microsecond response times and produce no photocurrent. We engineered a channelrhodopsin actuator, CheRiff, which shows high light sensitivity and rapid kinetics and is spectrally orthogonal to the QuasArs. A coexpression vector, Optopatch, enabled cross-talk-free genetically targeted all-optical electrophysiology. In cultured rat neurons, we combined Optopatch with patterned optical excitation to probe back-propagating action potentials (APs) in dendritic spines, synaptic transmission, subcellular microsecond-timescale details of AP propagation, and simultaneous firing of many neurons in a network. Optopatch measurements revealed homeostatic tuning of intrinsic excitability in human stem cell-derived neurons. In rat brain slices, Optopatch induced and reported APs and subthreshold events with high signal-to-noise ratios. The Optopatch platform enables high-throughput, spatially resolved electrophysiology without the use of conventional electrodes.
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JCR | SNIP | JCR | SNIP | JCR | SNIP | JCR | SNIP |
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in: SICRIS
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