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Imaging Membrane Potential with Two Types of Genetically Encoded Fluorescent Voltage Sensors
Published on: February 4, 2016
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A high-speed, bright, red fluorescent voltage sensor to detect neural activity
1Department of Biomedical Engineering, Duke University, Durham, NC, 27708, USA.
Scientific Reports
|November 6, 2019
Summary
Researchers developed Ace-mScarlet, a novel red fluorescent genetically encoded voltage indicator (GEVI). This advanced GEVI offers high sensitivity and brightness for optical neural recording, overcoming limitations of existing red sensors.
Area of Science:
- Neuroscience
- Biotechnology
- Optical Imaging
Background:
- Genetically encoded voltage indicators (GEVIs) enable optical recording of neural activity with high temporal resolution.
- Existing red-fluorescent GEVIs lack the performance and spectral separation of green fluorescent protein-based sensors.
- Challenges include limited response, brightness, and spectral overlap with other optical tools.
Purpose of the Study:
- To develop a red-fluorescent GEVI with improved performance and spectral characteristics.
- To overcome the limitations of current red-fluorescent voltage indicators.
- To enable simultaneous optical recording and optogenetic manipulation.
Main Methods:
- Fusion of Ace2N (voltage-sensitive rhodopsin) with mScarlet (red fluorescent protein) to create Ace-mScarlet.
- Utilizing Förster Resonance Energy Transfer (FRET) for voltage sensing.
- Characterization of spectral properties and performance in voltage detection.
Main Results:
- Ace-mScarlet exhibits high sensitivity, brightness, and fast kinetics comparable to green sensors.
- Its red-shifted spectra allow for excellent spectral separation from green sensors and blue-light actuators.
- Demonstrated high-fidelity voltage recordings during simultaneous optogenetic perturbation.
Conclusions:
- Ace-mScarlet is a powerful new red-fluorescent GEVI.
- It overcomes spectral limitations, enabling dual-color imaging and combined optical recording/perturbation.
- This technology advances neural activity monitoring and manipulation.
