Bright and fast multicoloured voltage reporters via electrochromic FRET

Peng Zou, Yongxin Zhao, Adam D. Douglass, Daniel R. Hochbaum, Daan Brinks, Christopher A. Werley, D. Jed Harrison, Robert E. Campbell () and Adam E. Cohen ()
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Peng Zou: Harvard Stem Cell Institute, Harvard University
Yongxin Zhao: University of Alberta
Adam D. Douglass: University of Utah
Daniel R. Hochbaum: Harvard Stem Cell Institute, Harvard University
Daan Brinks: Harvard Stem Cell Institute, Harvard University
Christopher A. Werley: Harvard Stem Cell Institute, Harvard University
D. Jed Harrison: University of Alberta
Robert E. Campbell: University of Alberta
Adam E. Cohen: Harvard Stem Cell Institute, Harvard University

Nature Communications, 2014, vol. 5, issue 1, 1-10

Abstract: Abstract Genetically encoded fluorescent reporters of membrane potential promise to reveal aspects of neural function not detectable by other means. We present a palette of multicoloured brightly fluorescent genetically encoded voltage indicators with sensitivities from 8–13% ΔF/F per 100 mV, and half-maximal response times from 4–7 ms. A fluorescent protein is fused to an archaerhodopsin-derived voltage sensor. Voltage-induced shifts in the absorption spectrum of the rhodopsin lead to voltage-dependent nonradiative quenching of the appended fluorescent protein. Through a library screen, we identify linkers and fluorescent protein combinations that report neuronal action potentials in cultured rat hippocampal neurons with a single-trial signal-to-noise ratio from 7 to 9 in a 1 kHz imaging bandwidth at modest illumination intensity. The freedom to choose a voltage indicator from an array of colours facilitates multicolour voltage imaging, as well as combination with other optical reporters and optogenetic actuators.

Date: 2014
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DOI: 10.1038/ncomms5625

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