TY - JOUR
T1 - Screening and Cellular Characterization of Genetically Encoded Voltage Indicators Based on Near-Infrared Fluorescent Proteins
AU - Monakhov, Mikhail V.
AU - Matlashov, Mikhail E.
AU - Colavita, Michelangelo
AU - Song, Chenchen
AU - Shcherbakova, Daria M.
AU - Antic, Srdjan D.
AU - Verkhusha, Vladislav V.
AU - Knöpfel, Thomas
N1 - Funding Information:
We thank all members of the Knöpfel, Verkhusha, and Antic laboratories for the useful discussions and suggestions. We thank Adam Cohen for the CheRiff spectra. This work was supported by the Grants NS099573, MH109091, NS103573, and GM122567 from the National Institutes of Health.
Publisher Copyright:
© 2020 American Chemical Society.
PY - 2020/11/4
Y1 - 2020/11/4
N2 - We developed genetically encoded voltage indicators using a transmembrane voltage-sensing domain and bright near-infrared fluorescent proteins derived from bacterial phytochromes. These new voltage indicators are excited by 640 nm light and emission is measured at 670 nm, allowing imaging in the near-infrared tissue transparency window. The spectral properties of our new indicators permit seamless voltage imaging with simultaneous blue-green light optogenetic actuator activation as well as simultaneous voltage-calcium imaging when paired with green calcium indicators. Iterative optimizations led to a fluorescent probe, here termed nirButterfly, which reliably reports neuronal activities including subthreshold membrane potential depolarization and hyperpolarization as well as spontaneous spiking or electrically- and optogenetically evoked action potentials. This enables largely improved all-optical causal interrogations of physiology.
AB - We developed genetically encoded voltage indicators using a transmembrane voltage-sensing domain and bright near-infrared fluorescent proteins derived from bacterial phytochromes. These new voltage indicators are excited by 640 nm light and emission is measured at 670 nm, allowing imaging in the near-infrared tissue transparency window. The spectral properties of our new indicators permit seamless voltage imaging with simultaneous blue-green light optogenetic actuator activation as well as simultaneous voltage-calcium imaging when paired with green calcium indicators. Iterative optimizations led to a fluorescent probe, here termed nirButterfly, which reliably reports neuronal activities including subthreshold membrane potential depolarization and hyperpolarization as well as spontaneous spiking or electrically- and optogenetically evoked action potentials. This enables largely improved all-optical causal interrogations of physiology.
KW - all-optical electrophysiology
KW - biosensor
KW - Butterfly
KW - FRET
KW - GEVI
KW - iRFP
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U2 - 10.1021/acschemneuro.0c00046
DO - 10.1021/acschemneuro.0c00046
M3 - Article
C2 - 33063984
AN - SCOPUS:85095671572
VL - 11
SP - 3523
EP - 3531
JO - ACS Chemical Neuroscience
JF - ACS Chemical Neuroscience
SN - 1948-7193
IS - 21
ER -