All-optical inter-layers functional connectivity investigation in the mouse retina

Spampinato, Giulia Lia Beatrice; Ronzitti, Emiliano; Zampini, Valeria; Trapani, Francesco; Khabou, Hanen; Agraval, Anurag; Dalkara, Deniz; Picaud, Serge; Papagiakoumou, Eirini; Marre, Olivier; Emiliani, Valentina

doi:10.1016/j.crmeth.2022.100268

Cited by 9 publications

(17 citation statements)

References 74 publications

(102 reference statements)

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“…A new study by Spampinato et al. (2022) introduces a powerful synthesis of approaches that may begin to elucidate the functional connectivity in many brain circuits.…”

Section: Main Textmentioning

confidence: 99%

“…In more detail, Spampinato et al. (2022) begin by virally expressing an optogenetic construct, CoChR ( Klapoetke et al., 2014 ), in a specific type of retinal bipolar cell called rod bipolar cells ( Figure 1 ).…”

Section: Main Textmentioning

confidence: 99%

“…Because bipolar cells generally signal using graded changes in their membrane potential, instead of action potentials, the authors also show that this holographic photostimulation technique can produce graded changes in the activation of bipolar cells.

Figure 1 Holographic photostimulation and GaCAMP imaging in ex vivo retina Spampinato et al. (2022) designed a system combining two-photon guided holographic photostimulation and calcium fluorescence imaging to study functional connectivity in ex vivo retina.…”

Section: Main Textmentioning

confidence: 99%

“… Spampinato et al. (2022) designed a system combining two-photon guided holographic photostimulation and calcium fluorescence imaging to study functional connectivity in ex vivo retina.…”

Section: Main Textmentioning

confidence: 99%

“…While certainly useful and interesting, behavior-based approaches do not reveal the functional connectivity among neurons. To meet this challenge, Spampinato et al. (2022) express a calcium-dependent fluorescence indicator (GCaMP6s) ( Chen et al., 2013 ) in RGCs while also expressing CoChR in rod bipolar cells.…”

Section: Main Textmentioning

confidence: 99%

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An optical approach for mapping functional connectivity at single-cell resolution in brain circuits

Roy

Field

2022

Cell Reports Methods

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“…A new study by Spampinato et al. (2022) introduces a powerful synthesis of approaches that may begin to elucidate the functional connectivity in many brain circuits.…”

Section: Main Textmentioning

confidence: 99%

Section: Main Textmentioning

confidence: 99%

Section: Main Textmentioning

confidence: 99%

“… Spampinato et al. (2022) designed a system combining two-photon guided holographic photostimulation and calcium fluorescence imaging to study functional connectivity in ex vivo retina.…”

Section: Main Textmentioning

confidence: 99%

Section: Main Textmentioning

confidence: 99%

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An optical approach for mapping functional connectivity at single-cell resolution in brain circuits

Roy

Field

2022

Cell Reports Methods

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Ultrafast light targeting for high-throughput precise control of neuronal networks

et al. 2023

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Two-photon, single-cell resolution optogenetics based on holographic light-targeting approaches enables the generation of precise spatiotemporal neuronal activity patterns and thus a broad range of experimental applications, such as high throughput connectivity mapping and probing neural codes for perception. Yet, current holographic approaches limit the resolution for tuning the relative spiking time of distinct cells to a few milliseconds, and the achievable number of targets to 100-200, depending on the working depth. To overcome these limitations and expand the capabilities of single-cell optogenetics, we introduce an ultra-fast sequential light targeting (FLiT) optical configuration based on the rapid switching of a temporally focused beam between holograms at kHz rates. We used FLiT to demonstrate two illumination protocols, termed hybrid- and cyclic-illumination, and achieve sub-millisecond control of sequential neuronal activation and high throughput multicell illumination in vitro (mouse organotypic and acute brain slices) and in vivo (zebrafish larvae and mice), while minimizing light-induced thermal rise. These approaches will be important for experiments that require rapid and precise cell stimulation with defined spatio-temporal activity patterns and optical control of large neuronal ensembles.

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Scanless two-photon voltage imaging

Sims,

Bendifallah,

Grimm

et al. 2024

Nat Commun

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Two-photon voltage imaging has long been heralded as a transformative approach capable of answering many long-standing questions in modern neuroscience. However, exploiting its full potential requires the development of novel imaging approaches well suited to the photophysical properties of genetically encoded voltage indicators. We demonstrate that parallel excitation approaches developed for scanless two-photon photostimulation enable high-SNR two-photon voltage imaging. We use whole-cell patch-clamp electrophysiology to perform a thorough characterization of scanless two-photon voltage imaging using three parallel illumination approaches and lasers with different repetition rates and wavelengths. We demonstrate voltage recordings of high-frequency spike trains and sub-threshold depolarizations from neurons expressing the soma-targeted genetically encoded voltage indicator JEDI-2P-Kv. Using a low repetition-rate laser, we perform multi-cell recordings from up to fifteen targets simultaneously. We co-express JEDI-2P-Kv and the channelrhodopsin ChroME-ST and capitalize on their overlapping two-photon absorption spectra to simultaneously evoke and image action potentials using a single laser source. We also demonstrate in vivo scanless two-photon imaging of multiple cells simultaneously up to 250 µm deep in the barrel cortex of head-fixed, anaesthetised mice.

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All-optical inter-layers functional connectivity investigation in the mouse retina

Cited by 9 publications

References 74 publications

An optical approach for mapping functional connectivity at single-cell resolution in brain circuits

An optical approach for mapping functional connectivity at single-cell resolution in brain circuits

Ultrafast light targeting for high-throughput precise control of neuronal networks

Scanless two-photon voltage imaging

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