Two-Photon Optogenetics by Computer-Generated Holography

Papagiakoumou, Eirini; Ronzitti, Emiliano; Chen, I‐Wen; Gajowa, Marta; Picot, Alexis; Emiliani, Valentina

doi:10.1007/978-1-4939-7417-7_10

Cited by 16 publications

(12 citation statements)

References 74 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The three opsins differ in their photocycle properties related to channel opening, closing, and inactivating (Nagel et al, 2003;Nikolic et al, 2009), which may play important roles in the temporal properties of 2P activation. ReaChR is a slow opsin, displaying the longest transition time spans between photocycle states (off-time constant: ϳ90 -150 ms; Lin et al, 2013;Chaigneau et al, 2016;Papagiakoumou et al, 2018). Meanwhile, ChrimsonR is the fastest of the three opsins (off-time constant: ϳ15 ms).…”

Section: Introductionmentioning

confidence: 99%

In vivo sub-millisecond two-photon optogenetics with temporally focused patterned light

et al. 2019

Self Cite

View full text Add to dashboard Cite

To better examine circuit mechanisms underlying perception and behavior, researchers need tools to enable temporally precise control of action-potential generation of individual cells from neuronal ensembles. Here we demonstrate that such precision can be achieved with two-photon (2P) temporally focused computer-generated holography to control neuronal excitability at the supragranular layers of anesthetized and awake visual cortex in both male and female mice. Using 2P-guided whole-cell or cell-attached recordings in positive neurons expressing any of the three opsins ReaChR, CoChR, or ChrimsonR, we investigated the dependence of spiking activity on the opsin's channel kinetics. We found that in all cases the use of brief illumination (Յ10 ms) induces spikes of millisecond temporal resolution and submillisecond precision, which were preserved upon repetitive illuminations up to tens of hertz. To reach high temporal precision, we used a large illumination spot covering the entire cell body and an amplified laser at high peak power and low excitation intensity (on average Յ0.2 mW/m 2 ), thus minimizing the risk for nonlinear photodamage effects. Finally, by combining 2P holographic excitation with electrophysiological recordings and calcium imaging using GCaMP6s, we investigated the factors, including illumination shape and intensity, opsin distribution in the target cell, and cell morphology, which affect the spatial selectivity of single-cell and multicell holographic activation. Parallel optical control of neuronal activity with cellular resolution and millisecond temporal precision should make it easier to investigate neuronal connections and find further links between connectivity, microcircuit dynamics, and brain functions.Recent developments in the field of optogenetics has enabled researchers to probe the neuronal microcircuit with light by optically actuating genetically encoded light-sensitive opsins expressed in the target cells. Here, we applied holographic light shaping and temporal focusing to simultaneously deliver axially confined holographic patterns to opsin-positive cells in the living mouse cortex. Parallel illumination efficiently induced action potentials with high temporal resolution and precision for three opsins of different kinetics. We extended the parallel optogenetic activation at low intensity to multiple neurons and concurrently monitored their calcium dynamics. These results demonstrate fast and temporally precise in vivo control of a neuronal subpopulation, opening new opportunities for revealing circuit mechanisms underlying brain functions. Chen, Ronzitti et al. • In vivo 2P Holographic Optogenetics

show abstract

Section: Introductionmentioning

confidence: 99%

In vivo sub-millisecond two-photon optogenetics with temporally focused patterned light

et al. 2019

Self Cite

View full text Add to dashboard Cite

show abstract

“…With this approach, it was possible to drive tail bending by selective photoactivation of specific ensemble of premotor neurons in the larval zebrafish brain 35 . Similarly to the case of 2D-CGH, illumination of spatially closed targets quickly deteriorates the axial resolution 36 . On the other hand using 3D illumination with TF is a challenge because the axially shifted holographic planes cannot be simultaneously imaged on the TF grating.…”

Section: Light Delivering Approachesmentioning

confidence: 99%

“…Overall τ on and τ inact have a non-linear dependence on light irradiance and depend on the excitation wavelength, while τ off can be considered independent of light irradiance. Notably, the kinetics parameters can largely differ from one opsin to another (Figure 3b) 36,46,50,55 . Fast opsins, such as Chronos, have τ on (at saturation) ≈1–2 ms and τ off ≈ 4 ms 36,50,55 , while slow opsins, such as ReaChR or C1V1 TT have τ on ≈ 6–8 ms (at saturation) and τ off ≈ 50–100 ms 45,50,56 .…”

Section: Opsin Engineeringmentioning

confidence: 99%

“…Notably, the kinetics parameters can largely differ from one opsin to another (Figure 3b) 36,46,50,55 . Fast opsins, such as Chronos, have τ on (at saturation) ≈1–2 ms and τ off ≈ 4 ms 36,50,55 , while slow opsins, such as ReaChR or C1V1 TT have τ on ≈ 6–8 ms (at saturation) and τ off ≈ 50–100 ms 45,50,56 . CoChR and ChrimsonR have intermediate values: τ on ≈ 2–6 ms, τ off ≈ 30 ms 46,50 , and τ on ≈ 8 ms, τ off ≈ 15 ms 50 , respectively.…”

Section: Opsin Engineeringmentioning

confidence: 99%

See 1 more Smart Citation

Towards circuit optogenetics

Chen

Papagiakoumou

Emiliani

2018

Current Opinion in Neurobiology

Self Cite

View full text Add to dashboard Cite

Optogenetics neuronal targeting combined with single-photon wide-field illumination has already proved its enormous potential in neuroscience, enabling the optical control of entire neuronal networks and disentangling their role in the control of specific behaviors. However, establishing how a single or a sub-set of neurons controls a specific behavior, or how functionally identical neurons are connected in a particular task, or yet how behaviors can be modified in real-time by the complex wiring diagram of neuronal connections requires more sophisticated approaches enabling to drive neuronal circuits activity with single-cell precision and millisecond temporal resolution. This has motivated on one side the development of flexible optical methods for two-photon (2P) optogenetic activation using either, or a hybrid of two approaches: scanning and parallel illumination. On the other side, it has stimulated the engineering of new opsins with modified spectral characteristics, channel kinetics and spatial distribution of expression, offering the necessary flexibility of choosing the appropriate opsin for each application. The need for optical manipulation of multiple targets with millisecond temporal resolution has imposed three-dimension (3D) parallel holographic illumination as the technique of choice for optical control of neuronal circuits organized in 3D. Today 3D parallel illumination exists in several complementary variants, each with a different degree of simplicity, light uniformity, temporal precision and axial resolution. In parallel, the possibility to reach hundreds of targets in 3D volumes has prompted the development of low-repetition rate amplified laser sources enabling high peak power, while keeping low average power for stimulating each cell. All together those progresses open the way for a precise optical manipulation of neuronal circuits with unprecedented precision and flexibility.

show abstract

“…The development of 3D photoactivation methods is more recent. These systems are based on the use of Computer-Generated Holography (CGH) (Papagiakoumou et al, 2018; Yang and Yuste, 2018). Although this technique was established for the projection of 3D patterns (Piestun et al, 1996; Haist et al, 1997) or diffraction-limited spots (Liesener et al, 2000) via spatial light modulators (SLMs) several years ago, its use in neuroscience for simultaneous activation of multiple targets in two (Lutz et al, 2008; Nikolenko et al, 2008; Dal Maschio et al, 2010) or three dimensions (Yang et al, 2011; Go et al, 2012; Hernandez et al, 2016; Dal Maschio et al, 2017) was established only during the last decade.…”

Section: Introductionmentioning

confidence: 99%

Methods for Three-Dimensional All-Optical Manipulation of Neural Circuits

Ronzitti

Emiliani

Papagiakoumou

2018

Front. Cell. Neurosci.

Self Cite

View full text Add to dashboard Cite

Optical means for modulating and monitoring neuronal activity, have provided substantial insights to neurophysiology and toward our understanding of how the brain works. Optogenetic actuators, calcium or voltage imaging probes and other molecular tools, combined with advanced microscopies have allowed an “all-optical” readout and modulation of neural circuits. Completion of this remarkable work is evolving toward a three-dimensional (3D) manipulation of neural ensembles at a high spatiotemporal resolution. Recently, original optical methods have been proposed for both activating and monitoring neurons in a 3D space, mainly through optogenetic compounds. Here, we review these methods and anticipate possible combinations among them.

show abstract

Two-Photon Optogenetics by Computer-Generated Holography

Cited by 16 publications

References 74 publications

In vivo sub-millisecond two-photon optogenetics with temporally focused patterned light

In vivo sub-millisecond two-photon optogenetics with temporally focused patterned light

Towards circuit optogenetics

Methods for Three-Dimensional All-Optical Manipulation of Neural Circuits

Contact Info

Product

Resources

About