2 articles published in JoVE
Laser Nanosurgery of Cerebellar Axons In Vivo Anna L. Allegra Mascaro1, Leonardo Sacconi1,2, Francesco Saverio Pavone1,2,3,4 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.
Micron-scale Resolution Optical Tomography of Entire Mouse Brains with Confocal Light Sheet Microscopy Ludovico Silvestri1, Alessandro Bria2,3, Irene Costantini1, Leonardo Sacconi1,4, Hanchuan Peng5, Giulio Iannello2, Francesco Saverio Pavone1,4,6,7 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.