26. February 2019

Francesco S. Pavone

Lagere area functional and structural nonlinear brain imaging

LENS-University of Florence / Sesto Fiorentino, Italy

High-resolution microscopy methods based on nonlinear optical technique find numerous applications in brain imaging. Functional or structural information can be gain with different implementations. In this work large area reconstruction are obtained using a mesoscale two-photon system for structural analysis and a light sheet two-photon microscope for functional information. The mesoscale methodology developed allows analyzing the cytoarchitecture of the human brain in three dimensions at high resolution. The combination of experimental protocols, based on optical tissue clearing and autofluorescence enhancement, with an automated software analysis enable to expand the histological studies to the third dimension. Functional imaging has been used to investigate whole organ, like zebrafish larval brain activity, using standard scanning or light sheet two-photon illumination. Both modalities are capable to sample whole brain with single cell resolution, with light sheet imaging being capable to perform high rate volumetric imaging allowing to map in real time whole-brain calcium dynamics not affected by undesired visual stimulation artefacts, as occurring in one photon excitation fluorescence microscopy. Large area non linear imaging allows in general extended measurements of the neuronal activity in normal conditions (spontaneous activity), under pathological modeling of epilepsy (seizure activity) and during visual stimulation (sensorial stimulated activity). 

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