Dr. Francesco A. Resta Profile

Dr. Francesco A. Resta

at LENS - Lab Europeo di Spettroscopie Non-Lineari

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Publications (7)

Proceedings Article | 30 May 2022 Presentation

All-optical perturbational approach to study information integration among distributed cortical regions

Francesco Resta, Giacomo Mazzamuto, Emilia Conti, Anna Letizia Allegra Mascaro, Francesco Saverio Pavone

Proceedings Volume PC12144, PC1214405 (2022) https://doi.org/10.1117/12.2621619

KEYWORDS: Optogenetics, Information fusion, Consciousness, Brain, Luminescence, Imaging systems, Calcium, Visualization, Temporal resolution, Synapsins

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Brain responsiveness and its activation complexity are linked to the level of consciousness (Tononi et al., 2004). However, how these features change across brain states is still not clear. The combination of Transcranial Magnetic Stimulation and hd-EEG recordings represents the standard method to address this issue in humans. A preclinical analogous in lab animals would provide novel mechanistic insights on the brain-state-dependent complexity of the brain. A powerful technique to study mesoscale cortical connectivity in mice exploits wide-field fluorescence microscopy. This approach provides simultaneous information of neuronal ensemble activity from distributed cortical areas, while optogenetic has been demonstrated to be a powerful tool to activate cortical neuronal clusters. However, all-optical systems that combine these techniques critically suffer from crosstalk between imaging and photostimulation (Emiliani et al. 2015). Brain responsiveness and its activation complexity are linked to the level of consciousness (Tononi et al., 2004). However, how these features change across brain states is still not clear. The combination of Transcranial Magnetic Stimulation and hd-EEG recordings represents the standard method to address this issue in humans. A preclinical analogous in lab animals would provide novel mechanistic insights on the brain-state-dependent complexity of the brain. A powerful technique to study mesoscale cortical connectivity in mice exploits wide-field fluorescence microscopy. This approach provides simultaneous information of neuronal ensemble activity from distributed cortical areas, while optogenetic has been demonstrated to be a powerful tool to activate cortical neuronal clusters. However, all-optical systems that combine these techniques critically suffer for crosstalk between imaging and photostimulation (Emiliani et al. 2015). Here we established an all-optical method combining wide-field fluorescence imaging of the red-shifted calcium indicator jRCaMP1b and transcranial optogenetic stimulation of Channelrhodopsin-2 (ChR2). To achieve a cortex-wide expression of the calcium indicator, an adeno-associated virus (AAV.PHP.eb) carrying jRCaMP1b under the control of the synapsin promoter was injected in the retro-orbital sinus of anesthetized mice. This led to a uniform expression of the functional indicator in the whole cortex, giving the possibility to visualize the neuronal activity propagation in all the cortical areas. Due to the high opsins expression required for effective optogenetic stimulation, AAV9-ChR2 was locally injected in the somatosensory cortex (S1). Results show that in awake mice, optogenetic stimulations at increasing laser power evoke a distributed cortical response in several areas in the two cortical hemispheres, whereas, during anesthesia, stimulation led to a localized reponse limited in space and time. These results suggest that response complexity decrease with the levels of consciousness, as observed in pathological patients affected by disorders of consciousness (Massimini et al., 2009).

Proceedings Article | 30 May 2022 Presentation

An AOD breakthrough for volumetric 2P optogenetic applications

Pietro Ricci, Marco Marchetti, Michele Sorelli, Lapo Turrini, Francesco Resta, Vladislav Gavryusev, Giuseppe de Vito, Giuseppe Sancataldo, Francesco Vanzi, Ludovico Silvestri, Francesco Pavone

Proceedings Volume PC12144, PC1214406 (2022) https://doi.org/10.1117/12.2620313

KEYWORDS: Optogenetics, Photostimulation, Brain, Spatial light modulators, Optical design, Neurons, Mirrors, Pathology, Microscopy, Laser scanners

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To understand the brain computation paradigms and the causal interactions in complex neuronal networks, we need methods and technologies to record and perturb neuronal distributions over large fields of view. In this application, two-photon (2P) imaging has become a cornerstone microscopy technique, widely used for deep optical access in biological samples and selective light targeting with submicrometric resolution. In parallel to structural and functional imaging, 2P optogenetics has represented a game-changer, allowing targeted stimulation of specific neural circuits. However, the long commutation times and refresh rates of traditional scanning methods substantially hinder near-simultaneous multi-site 3D stimulation. Acousto-optic deflectors (AODs), owing to their fastest scanning and refresh rates, can fulfil the temporal requirements for concurrent activation of sparsely distributed neurons. Nevertheless, their applicability to 2P optogenetics in large volumes has been limited so far by the massive efficiency drop along the optical axis during their use in axial scanning. To counteract this drawback, a compensation software module is frequently employed to flatten the power distribution throughout the volume. However, the power threshold is reduced to the minimum intensity value addressable, lowering the peak intensity released in the centre of the axial scan. Here, we propose a unique approach for overcoming this drawback which provided lifted axial power distribution while maintaining a uniform lateral illumination range. We tested this method by the 2P photoactivation of optogenetic actuators in 3D in zebrafish larvae, showing how the probability of evoking an electrophysiological response and the relative neuronal activity amplitude improved by carefully optimizing the light targeting time on different axial planes. In conclusion, fast and uniform axial light addressing with AODs enables unprecedented 3D 2P optostimulation, formerly not feasible. Furthermore, this approach can be adopted as an upgrade for existing microscopes designed for volumetric imaging, providing 3D multi-site imaging and random-access illumination.

Proceedings Article | 1 April 2020 Presentation

Movement-specific patterns of cortical activation revelated by in vivo all-optical imaging and manipulation of neuronal activity in the motor cortex (Conference Presentation)

Elena Montagni, Francesco Resta, Giuseppe de Vito, Alessandro Scaglione, Anna Letizia Allegra Mascaro, Francesco Saverio Pavone

Proceedings Volume 11360, 1136007 (2020) https://doi.org/10.1117/12.2560336

KEYWORDS: In vivo imaging, Optogenetics, Cortical activation, Calcium, Brain mapping, Organisms, Photostimulation, Radiofrequency ablation, Actuators, Control systems

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Proceedings Article | 21 February 2020 Presentation + Paper

Mesoscale imaging of neuronal activity coupled with light-evoked motor mapping reveal movement-specific spatiotemporal patterns of cortical activation in awake mice

Francesco Resta, Anna Letizia Allegra Mascaro, Elena Montagni, Giuseppe de Vito, Alessandro Scaglione, Francesco Saverio Pavone

Proceedings Volume 11226, 112260I (2020) https://doi.org/10.1117/12.2544214

KEYWORDS: Calcium, Brain mapping, Cortical activation, Optogenetics, Imaging systems, Neuroscience, Neurons, Light sources

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Proceedings Article | 22 July 2019 Paper

Full-optical stimulation and readout of neuronal activity during optogenetically-evoked movements in awake mice

Francesco Resta, Elena Montagni, Giuseppe De Vito, Alessandro Scaglione, Anna Letizia Allegra Mascaro, Francesco Saverio Pavone

Proceedings Volume 11076, 1107609 (2019) https://doi.org/10.1117/12.2527112

KEYWORDS: Calcium, Optogenetics, Pulsed laser operation, Astronomical imaging, Actuators, Brain, Statistical analysis, Nonlinear optics, Spectroscopes, Physics

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Proceedings Article | 4 March 2019 Presentation

All-optical readout and stimulation of cortical activity during optogenetically-triggered motor task in awake mice (Conference Presentation)

Francesco Resta, Elena Montagni, Emilia Conti, Giuseppe de Vito, Anna Letizia Allegra Mascaro, Francesco Saverio Pavone

Proceedings Volume 10865, 108650Y (2019) https://doi.org/10.1117/12.2508322

KEYWORDS: Radiofrequency ablation, Optogenetics, Calcium, Pulsed laser operation, Cortical activation

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Proceedings Article | 1 March 2019 Paper

Combined rehabilitation promotes recovery of motor functionality in a mouse model of stroke

Emilia Conti, Anna Letizia Allegra Mascaro, Francesco Resta, Alessandro Scaglione, Maria Pasquini, Silvestro Micera, Francesco Pavone

Proceedings Volume 10865, 108650R (2019) https://doi.org/10.1117/12.2508321

KEYWORDS: Optogenetics, Neurons, Calcium, Robotics, Skull, Mouse models, Therapeutics, Light emitting diodes, Cameras

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Showing 5 of 7 publications

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