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Allen Institute for Brain Science

3 ARTICLES PUBLISHED IN JoVE

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Neuroscience

Micron-scale Resolution Optical Tomography of Entire Mouse Brains with Confocal Light Sheet Microscopy
Ludovico Silvestri 1, Alessandro Bria 2,3, Irene Costantini 1, Leonardo Sacconi 1,4, Hanchuan Peng 5, Giulio Iannello 2, 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.

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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 Chong 1, Gilberto Soler-Llavina 1, Charles Cobbs 2, Christof Koch 1, Hongkui Zeng 1, Ed Lein 1
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.

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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.

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