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All-optical crosstalk-free manipulation and readout of Chronos-expressing Neurons

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Title: All-optical crosstalk-free manipulation and readout of Chronos-expressing Neurons
Authors: Soor, N
Quicke, P
Howe, C
Pang, KT
Neil, M
Schultz, S
Foust, A
Item Type: Journal Article
Abstract: All optical neurophysiology allows manipulation and readout of neural network activity with single-cell spatial resolution and millisecond temporal resolution. Neurons can be made to express proteins that actuate transmembrane currents upon light absorption, enabling optical control of membrane potential and action potential signalling. In addition, neurons can be genetically or synthetically labelled with fluorescent reporters of changes in intracellular calcium concentration or membrane potential. Thus, to optically manipulate and readout neural activity in parallel, two spectra are involved: the action spectrum of the actuator, and the absorption spectrum of the fluorescent reporter. Due to overlap in these spectra, previous all-optical neurophysiology paradigms have been hindered by spurious activation of neuronal activity caused by the readout light. Here, we pair the blue-green absorbing optogenetic actuator, Chronos, with a deep red-emitting fluorescent calcium reporter CaSiR-1. We show that cultured Chinese hamster ovary cells transfected with Chronos do not exhibit transmembrane currents when illuminated with wavelengths and intensities suitable for exciting one-photon CaSiR-1 fluorescence. We then demonstrate crosstalk-free, high signal-to-noise ratio CaSiR-1 red fluorescence imaging at 100 frames s−1 of Chronos-mediated calcium transients evoked in neurons with blue light pulses at rates up to 20 Hz. These results indicate that the spectral separation between red light excited fluorophores, excited efficiently at or above 640 nm, with blue-green absorbing opsins such as Chronos, is sufficient to avoid spurious opsin actuation by the imaging wavelengths and therefore enable crosstalk-free all-optical neuronal manipulation and readout.
Issue Date: 9-Jan-2019
Date of Acceptance: 13-Dec-2018
URI: http://hdl.handle.net/10044/1/66803
DOI: https://dx.doi.org/10.1088/1361-6463/aaf944
ISSN: 0022-3727
Publisher: IOP Publishing
Journal / Book Title: Journal of Physics D: Applied Physics
Volume: 52
Issue: 10
Copyright Statement: © 2019 IOP Publishing Ltd. Original content from this work may be used under the terms of the Creative Commons Attribution 3.0 licence. Any further distribution of this work must maintain attribution to the author(s) and the title of the work, journal citation and DOI
Sponsor/Funder: Biotechnology and Biological Sciences Research Council (BBSRC)
Engineering & Physical Science Research Council (E
Royal Academy Of Engineering
National Institutes of Health
Wellcome Trust
Biotechnology and Biological Sciences Research Council (BBSRC)
Funder's Grant Number: BB/K001817/1
EP/K503733/1
RF1415\14\26
UPMC: C15/0244
201964/Z/16/Z
BB/R009007/1
Keywords: Science & Technology
Physical Sciences
Physics, Applied
Physics
Chronos
calcium indicator
optogenetics
crosstalk
functional imaging
neuron
neuromodulation
TRANSGENIC MICE
RED
OPTOGENETICS
EXCITATION
DYE
CHANNELRHODOPSIN-2
ELECTROPHYSIOLOGY
INTERROGATION
INHIBITION
INDICATORS
02 Physical Sciences
09 Engineering
Applied Physics
Publication Status: Published
Article Number: 104002
Online Publication Date: 2019-01-09
Appears in Collections:Physics
Bioengineering
Photonics
Faculty of Natural Sciences



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