Dr. Vladislav Gavryusev Profile

Dr. Vladislav Gavryusev

at Univ degli Studi di Firenze

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Publications (6)

Proceedings Article | 16 May 2023 Presentation + Paper

High-speed light-sheet microscopy imaging and data post-processing using custom hardware and software solutions

Giacomo Mazzamuto, Ludovico Silvestri, Giuseppe Sancataldo, Vladislav Gavryusev, Marina Scardigli, Irene Costantini, Francesco Pavone

Proceedings Volume 12385, 1238504 (2023) https://doi.org/10.1117/12.2650374

KEYWORDS: Brain, Data acquisition, Cameras, Microscopy, Microscopes, Open source software, Image compression, Equipment, Acoustooptic tunable filters, Biological samples

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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

Confocal detection without striping artifacts exploiting a fast multidirectional DSLM (Conference Presentation)

Pietro Ricci, Giuseppe Sancataldo, Alessandra Franceschini, Vladislav Gavryusev, Ludovico Silvestri, Francesco Saverio Pavone

Proceedings Volume 11360, 1136006 (2020) https://doi.org/10.1117/12.2560261

KEYWORDS: Confocal microscopy, Microscopy, Luminescence, Signal detection, Light scattering, Neuroscience, Stereoscopy, Image acquisition, Image processing, Absorption

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In the past few years, Light Sheet Fluorescence Microscopy (LSFM) has become a cornerstone imaging technique for neuroscience, improving the quality and capabilities of 3D imaging. Selec-tive illumination of a single plane provides intrinsic optical sectioning and fast image recording, while minimizing out-of-focus fluorescence background, sample photo-damage and photobleach-ing. However, images acquired with LSFM are often affected by light absorption or scattering effects, leading to un-even illumination and striping artifacts. Here we present an optical solution for such problem, via fast multi-directional illumination of the sample, based on an Acousto-Optical Deflector (AOD). Using this device, we were able to pivot the beam respect the propagation axis, with a scanning rate faster than the detector acquisition rate, in order to average the shadows attenuation at different angles over time. We also demonstrate that this scanning AOD system is compatible with Digital Scanned laser Light-sheet fluorescence microscopy (DSLM). Furthermore, we provided a theoretical model of the beam pivoting, looking for the optimal beam parameters to optimize the detection efficiency. It has been done because we wanted to adapt such scanning technique to Confocal Light Sheet Microscopy (CLSM), which intrinsically shows an incompatibility between the beam pivoting and the finite size of a digital slit used to spatially filter out spurious signals. We partially solved the problem expanding the beam and using two cylindrical lenses to create an elliptical-Gaussian beam which enables to cover more the digital slit while pivoting the beam. We test its performance by acquiring several mouse brain areas, observing real-time shadows suppression while preserving confocal detection of the signal emitted by specific fluorophores. A comparison between such scanning beam illumination and a traditional static one has been carried out in term of contrast analysis, striping suppression and Point Spread Function response.

Proceedings Article | 1 April 2020 Presentation + Paper

Fast volumetric mapping of human brain slices

Luca Pesce, Annunziatina Laurino, Vladislav Gavryusev, Giacomo Mazzamuto, Giuseppe Sancataldo, Marina Scardigli, Matteo Roffilli, Ludovico Silvestri, Irene Costantini, Francesco Saverio Pavone

Proceedings Volume 11360, 113600A (2020) https://doi.org/10.1117/12.2555644

KEYWORDS: Brain, Neuroimaging, Microscopy, Tissues, Luminescence, Brain mapping, Neurons, Refractive index, Switches, Tissue optics

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Proceedings Article | 21 February 2020 Presentation + Paper

Fast volumetric mapping of human brain slices

Luca Pesce, Annunziatina Laurino, Vladislav Gavryusev, Giacomo Mazzamuto, Giuseppe Sancataldo, Marina Scardigli, Matteo Roffilli, Ludovico Silvestri, Irene Costantini, Francesco Saverio Pavone

Proceedings Volume 11226, 112260B (2020) https://doi.org/10.1117/12.2544133

KEYWORDS: Brain, Brain mapping, Neuroimaging, Tissues, Refractive index, Microscopy, Neurons, Tissue optics, Tissue clearing

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SPIE Journal Paper | 31 October 2019 Open Access

Dual-beam confocal light-sheet microscopy via flexible acousto-optic deflector

Vladislav Gavryusev, Giuseppe Sancataldo, Pietro Ricci, Alberto Montalbano, Chiara Fornetto, Lapo Turrini, Annunziatina Laurino, Luca Pesce, Giuseppe de Vito, Natascia Tiso, Francesco Vanzi, Ludovico Silvestri, Francesco Pavone

JBO, Vol. 24, Issue 10, 106504, (October 2019) https://doi.org/10.1117/12.10.1117/1.JBO.24.10.106504

KEYWORDS: Confocal microscopy, Camera shutters, Cameras, Microscopy, Brain, Acousto-optics, Sensors, Luminescence, Neuroimaging, Image acquisition

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