In Adam et al. 2018, a multispot two-photon excitation microscope with rolling shutter wide-field detection was developed. This method benefits from the increased penetration depth of two-photon excitation in scattering media and the increased imaging rates of multiplexed excitation while exploiting the rolling shutter of a fast sCMOS camera. Microscope performances were demonstrated in fixed semicleared brain slices by imaging dendritic spines up to 400-μm deep.
In Montagni et al. 2018 we describe the evaluation of different indicators of neuronal activity to identify the most suited for the development of an all-optical system. We tested different red-shifted genetically encoded calcium indicators to identify the best one to reveal the cortical neuronal activity and a combination of indicator and actuator that could be successfully used to read and write from mouse cortex, towards the intra and inter subject transfer of neuronal functionality. We identified jRCaMP1a as the best red-shifted functional indicator that does not show photoswitch, resulting the best choice for the future development of an all-optical system for interrogation of neuronal circuits.
In Sancataldo et al. (Front. Neuroanat. 2019), we described the use of acousto-optic deflectors (AODs) in light-sheet microscopy to mitigate striping artefacts. This work paves the way to a more quantitative functional imaging in zebrafish larvae, which is a key aspect of the project, and which was also previously investigated by our group (Muellenbroich et al., Front. Cell Neurosci. 2018).
Further, the use of AODs has been a technical training for the implementation of multiplexed ultrafast light-sheet microscopy.
In de Vito et al. (Biom. Opt. Express, 2022), we present a microscope that quintuplicated the previous maximal volumetric acquisition speed of a vertebrate brain (zebrafish larva) with cellular resolution. We used this microscope to characterize brain-wide events of fast neuronal activity (caudo-rostral ictal waves) that were previously unreported.
In Pisanello M. et al. (Sci. Rep., 2018), we showed how tapered optical fibers can be a versatile tool for performing light delivery in both shallow and deep brain areas. Moreover, we showed how it is possible to reconfigure and tune light emission patterns in order to be adapted to the brain region of interest, also dynamically switching from restricted or wide illumination of brain volumes.
In Pisano et al. (Nat. Meth., 2019), we described how the modal properties of the tapered optical fibers can also lead to the possibility of collecting light thus performing fiber photometry. In combination with fiber microstructuring and tailoring of the light collection volumes, we demonstrated in vivo fiber photometry of dopaminergic activity in the dorsal vs ventral striatum of behaving mice.
In Spagnolo B. et al. (Nat. Mater., 2022) we implemented microstructured optical windows and microelectrodes on the taper of the fiber by exploiting two-photon lithography in order to pattern different geometries of electrically and optically active elements all along and all around the taper axis. This approach greatly maximizes the possibility of studying deep brain circuitry with customized light-delivery patterns while recording neural activity, with no photoelectric artefacts thanks to the optimized microfabrication protocol.
In Quarta et al., 2022 we investigated the hypothesis that areas beyond the motor regions could participate in reach to grasp planning and execution. We found that a large network of areas is engaged while performing the reach to grasp task and the kinematics correlates mostly with neural activity in sensorimotor areas
In Allegra Mascaro et al., 2019, we investigate how rehabilitation paradigm affects neuronal and vascular plasticity of the mouse cortex after focal stroke. We fund that synaptic stabilization is associated with angiogenesis and recovery of a segregated motor representation.
At CNR we successfully consolidated the results obtained with high impact publications: Spalletti et al. 2017; Sammali et al., 2017; Alia et al., 2017, 2019, 2021; Conti et al., 2021.
