Optical studies of action potential dynamics with hVOS probes

Ma, Yihe; Bayguinov, Peter O.

doi:10.1016/j.cobme.2019.09.007

Cited by 8 publications

(8 citation statements)

References 45 publications

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“…These conformational changes are translated to the FP and change the fluorescence intensity of the FP. [203][204][205][206][207][208][209] GEVIs have been used to tackle intriguing scientific questions, such as how spiking activity of specific neuron types is modulated during different behavioral processes and relate to subthreshold membrane voltage fluctuations and extracellularly measured local field potential dynamics.…”

Section: Genetically Encoded Voltage Biosensorsmentioning

confidence: 99%

Neurophotonic Tools for Microscopic Measurements and Manipulation: Status Report

et al. 2022

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Section: Genetically Encoded Voltage Biosensorsmentioning

confidence: 99%

Neurophotonic Tools for Microscopic Measurements and Manipulation: Status Report

et al. 2022

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“…In particular, a few newly developed GEVIs, including SomArchon, QuasAr3, Voltron, ASAP3, and Ace2N, have been shown to be able to capture individual action potentials in behaving mice. Of these high-performance GEVIs, several are fully genetically encoded, whereas others are hybrid, requiring exogenous chemicals ( Bando et al., 2019 ; Beck et al., 2019 ; Kannan et al., 2019 ; Knöpfel and Song, 2019 ; Ma et al., 2019 ; Peng et al., 2017 ; Xu et al., 2017 ). One class of fully genetically encoded indicators utilizes fluorophores fused to voltage-sensitive protein domains derived from voltage-gated ion channels, voltage-sensitive phosphatases, or rhodopsins ( Bando et al., 2019 ; Beck et al., 2019 ; Kannan et al., 2019 ; Knöpfel and Song, 2019 ; Peng et al., 2017 ; Xu et al., 2017 ).…”

Section: Introductionmentioning

confidence: 99%

“…Another class of fully genetically encoded indicators is single compartment and detects the intrinsic voltage-dependent fluorescence of engineered rhodopsins, such as QuasAr3, Archon, and SomArchon (Adam et al, 2019;Piatkevich et al, 2018Piatkevich et al, , 2019. To improve fluorescence signals, bright chemical fluorophores have also been explored in the design of GEVIs, yielding a class of high-performance hybrid GEVIs that requires both exogeneous chemical dyes and the corresponding voltage-sensing protein counterparts (Abdelfattah et al, 2019(Abdelfattah et al, , 2020Ma et al, 2019). With rapid and continued improvements of GEVIs, voltage imaging offers an exciting opportunity for direct analysis of neuronal voltage in the mammalian brain.…”

Section: Introductionmentioning

confidence: 99%

Large-scale voltage imaging in behaving mice using targeted illumination

et al. 2021

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Summary Recent improvements in genetically encoded voltage indicators enabled optical imaging of action potentials and subthreshold transmembrane voltage in vivo . To perform high-speed voltage imaging of many neurons simultaneously over a large anatomical area, widefield microscopy remains an essential tool. However, the lack of optical sectioning makes widefield microscopy prone to background cross-contamination. We implemented a digital-micromirror-device-based targeted illumination strategy to restrict illumination to the cells of interest and quantified the resulting improvement both theoretically and experimentally with SomArchon expressing neurons. We found that targeted illumination increased SomArchon signal contrast, decreased photobleaching, and reduced background cross-contamination. With the use of a high-speed, large-area sCMOS camera, we routinely imaged tens of spiking neurons simultaneously over minutes in behaving mice. Thus, the targeted illumination strategy described here offers a simple solution for widefield voltage imaging of many neurons over a large field of view in behaving animals.

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“…In particular, a few recent GEVIs, including SomArchon, QuasAr3, Voltron, ASAP3 and Ace2N, have achieved sufficient sensitivity to capture individual action potentials from single neurons in behaving mice. Of these high performance GEVIs, several are fully genetically encoded, whereas others are hybrid sensors that require exogenous chemicals [8][9][10][11][12][13][14]17 . One class of fully genetically encoded indicators detects voltage dependent fluorescence of fluorophores fused to voltage sensitive peptide domains derived from voltage gated ion channels, voltage sensitive phosphatases or rhodopsins [8][9][10][11][12][13][14] .…”

Section: Introductionmentioning

confidence: 99%

“…Another class of fully genetically encoded indicators is single compartment and directly detects the intrinsic voltage dependent fluorescence of engineered rhodopsins, such as QuarsAR3, Archon and SomArchon 3,5,15 . To improve fluorescence signals, bright chemical fluorophores have also been explored in the design of GVEIs, yielding a class of high-performance hybrid GEVIs that requires both exogeneous chemical dyes and the corresponding voltage sensing protein counterparts 2,16,17 .…”

Section: Introductionmentioning

confidence: 99%

Large-scale voltage imaging in the brain using targeted illumination

Xiao

Lowet

Gritton

et al. 2021

Preprint

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Recent improvements in genetically encoded voltage indicators enabled high precision imaging of single neuron's action potentials and subthreshold membrane voltage dynamics in the mammalian brain. To perform high speed voltage imaging, widefield microscopy remains an essential tool to record activity from many neurons simultaneously over a large anatomical area. However, the lack of optical sectioning makes widefield microscopy prone to background signal contamination. We implemented a simple, low cost, targeted illumination strategy based on a digital micromirror device (DMD) to restrict illumination to the cells of interest to improve background rejection, and quantified optical voltage signal improvement in neurons expressing the fully genetically encoded voltage indicator SomArchon. We found that targeted illumination, in comparison to widefield illumination, increased SomArchon signal contrast and reduced background cross-contamination in the brains of awake mice. Such improvement permitted the reduction of illumination intensity, and thus reduced fluorescence photobleaching and prolonged imaging duration. When coupled with a high-speed sCMOS camera, we routinely imaged tens of spiking neurons simultaneously over several minutes in the brain. Thus, the DMD-based targeted illumination strategy described here offers a simple solution for high-speed voltage imaging analysis of large scale network at the millisecond time scale with single cell resolution in the brains of behaving animals.

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Optical studies of action potential dynamics with hVOS probes

Cited by 8 publications

References 45 publications

Neurophotonic Tools for Microscopic Measurements and Manipulation: Status Report

Neurophotonic Tools for Microscopic Measurements and Manipulation: Status Report

Large-scale voltage imaging in behaving mice using targeted illumination

Large-scale voltage imaging in the brain using targeted illumination

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