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Allen Institute for Brain Science

3 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

Preparation of Acute Brain Slices Using an Optimized N-Methyl-D-glucamine Protective Recovery Method

Jonathan T. Ting^*1, Brian R. Lee^*1, Peter Chong¹, Gilberto Soler-Llavina¹, Charles Cobbs², Christof Koch¹, Hongkui Zeng¹, Ed Lein¹

¹Cell Types Program, Allen Institute for Brain Science, ²The Ben and Catherine Ivy Center for Advanced Brain Tumor Treatment, Swedish Neuroscience Institute

This protocol demonstrates the implementation of an optimized N-methyl-D-glucamine (NMDG) protective recovery method of brain slice preparation. A single media formulation is used to reliably obtain healthy brain slices from animals of any age and for diverse experimental applications.

Neuroscience

Injections of AAV Vectors for Optogenetics in Anesthetized and Awake Behaving Non-Human Primate Brain

Yoshiko Kojima^1,2, Jonathan T. Ting^2,3,4, Robijanto Soetedjo^2,4, Shane D. Gibson^2,4, Gregory D. Horwitz^2,4

¹Dept. of Otolaryngology - Head and Neck Surgery, University of Washington, ²Washington National Primate Research Center, University of Washington, ³Allen Institute for Brain Science, ⁴Dept. of Physiology & Biophysics, University of Washington

As currently implemented, optogenetics in non-human primates requires injection of viral vectors into the brain. An optimal injection method should be reliable and, for many applications, capable of targeting individual sites of arbitrary depth that are readily and unambiguously identified in postmortem histology. An injection method with these properties is presented.