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Nat Commun. 2014 Aug 13;5:4625. doi: 10.1038/ncomms5625.

Bright and fast multicoloured voltage reporters via electrochromic FRET.

Author information

1
Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA 02138.
2
Department of Chemistry, University of Alberta, Edmonton, Alberta, Canada, T6G 2G2.
3
Department of Neurobiology and Anatomy, University of Utah, Salt Lake City, UT 84132.
4
Howard Hughes Medical Institute.
#
Contributed equally

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.

PMID:
25118186
PMCID:
PMC4134104
DOI:
10.1038/ncomms5625
[Indexed for MEDLINE]
Free PMC Article

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