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Curr Protoc Pharmacol. 2017 Sep 11;78:11.20.1-11.20.24. doi: 10.1002/cpph.25.

All-Optical Electrophysiology for Disease Modeling and Pharmacological Characterization of Neurons.

Author information

1
Q-State Biosciences, Cambridge, Massachusetts.

Abstract

A key challenge for establishing a phenotypic screen for neuronal excitability is measurement of membrane potential changes with high throughput and accuracy. Most approaches for probing excitability rely on low-throughput, invasive methods or lack cell-specific information. These limitations stimulated the development of novel strategies for characterizing the electrical properties of cultured neurons. Among these was the development of optogenetic technologies (Optopatch) that allow for stimulation and recording of membrane voltage signals from cultured neurons with single-cell sensitivity and millisecond temporal resolution. Neuronal activity is elicited using blue light activation of the channelrhodopsin variant 'CheRiff'. Action potentials and synaptic signals are measured with 'QuasAr', a rapid and sensitive voltage-indicating protein with near-infrared fluorescence that scales proportionately with transmembrane potential. This integrated technology of optical stimulation and recording of electrical signals enables investigation of neuronal electrical function with unprecedented scale and precision.

KEYWORDS:

CheRiff; Optical electrophysiology; Optopatch; QuasAr; disease modeling; induced pluripotent stem cell; optogenetics; voltage indicator

PMID:
28892145
PMCID:
PMC5721518
DOI:
10.1002/cpph.25
[Indexed for MEDLINE]
Free PMC Article

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