ICON Foundation, Sesto Fiorentino, Italy

2 ARTICLES PUBLISHED IN JoVE

Neuroscience

Micron-scale Resolution Optical Tomography of Entire Mouse Brains with Confocal Light Sheet Microscopy

Ludovico Silvestri¹, Alessandro Bria^2,3, Irene Costantini¹, Leonardo Sacconi^1,4, Hanchuan Peng⁵, Giulio Iannello², Francesco Saverio Pavone^1,4,6,7

¹European Laboratory for Non-linear Spectroscopy (LENS), ²Integrated Research Centre, University Campus Bio-medico of Rome, ³DAEMI, University of Cassino, ⁴National Institute of Optics (CNR-INO), ⁵Allen Institute for Brain Science, ⁶Department of Physics, University of Florence, ⁷ICON Foundation, Sesto Fiorentino, Italy

In this article we describe the full experimental procedure to reconstruct, with high resolution, the fine brain anatomy of fluorescently labeled mouse brains. The described protocol includes sample preparation and clearing, specimen mounting for imaging, data post-processing and multi-scale visualization.

Neuroscience

Laser Nanosurgery of Cerebellar Axons In Vivo

Anna L. Allegra Mascaro¹, Leonardo Sacconi^1,2, Francesco Saverio Pavone^1,2,3,4

¹European Laboratory for Non-Linear Spectroscopy, University of Florence, ²National Institute of Optics, National Research Council, ³Department of Physics and Astronomy, University of Florence, ⁴International Center for Computational Neurophotonics (ICON Foundation)

Two-photon imaging, coupled to laser nanodissection, are useful tools to study degenerative and regenerative processes in the central nervous system with subcellular resolution. This protocol shows how to label, image, and dissect single climbing fibers in the cerebellar cortex in vivo.