Dalhousie University View Institution's Website 9 articles published in JoVE Medicine Large-Animal Model of Donation after Circulatory Death and Normothermic Regional Perfusion for Cardiac Assessment Khalil Khalil*1,2, Roberto V. P. Ribeiro*3, Julgans S. Alvarez4,5, Mitesh V. Badiwala4,5, Shant Der Sarkissian1,2, Nicolas Noiseux1,2 1 The protocol describes a large-animal (porcine) model of donation after circulatory death, followed by thoracoabdominal normothermic regional perfusion that closely simulates the clinical scenario in heart transplantation, and has the potential to facilitate therapeutic studies and strategies. Immunology and Infection Intravital Widefield Fluorescence Microscopy of Pulmonary Microcirculation in Experimental Acute Lung Injury Using a Vacuum-Stabilized Imaging System Stefan Hall1, Sufyan Faridi2, Irene Euodia2,3, Sophie Tanner4, Andrew Krzysztof Chojnacki5, Kamala D. Patel6,7, Juan Zhou2,4, Christian Lehmann1,2,4,8 1Department of Physiology and Biophysics, Dalhousie University, 2Department of Microbiology and Immunology, Dalhousie University, 3Department of Neuroscience, Dalhousie University, 4Department of Anesthesia, Pain Management, and Perioperative Medicine, Dalhousie University, 5Live Cell Imaging Center, University of Calgary, 6Department of Physiology and Pharmacology, University of Calgary, 7Department of Biochemistry and Molecular Biology, University of Calgary, 8Department of Pharmacology, Dalhousie University Intravital fluorescence microscopy can be utilized to study leukocyte-endothelial interactions and capillary perfusion in real-time. This protocol describes methods to image and quantify these parameters in the pulmonary microcirculation using a vacuum-stabilized lung imaging system. Chemistry Organic Solvent-Based Protein Precipitation for Robust Proteome Purification Ahead of Mass Spectrometry Jessica L. Nickerson1, Venus Baghalabadi1, Ziheng Dang1, Victoria A. Miller2, Sara L. Little2, Alan A. Doucette1 1Department of Chemistry, Dalhousie University, 2Proteoform Scientific Inc. The present protocol describes solvent-based protein precipitation under controlled conditions for robust and rapid recovery and purification of proteome samples prior to mass spectrometry. Neuroscience A Standardized Pipeline for Examining Human Cerebellar Grey Matter Morphometry using Structural Magnetic Resonance Imaging Rebecca Kerestes1, Shuo Han2, Srinivas Balachander3, Carlos Hernandez-Castillo4, Jerry L. Prince5,6, Jörn Diedrichsen7, Ian H. Harding1,8 1Department of Neuroscience, Central Clinical School, Monash University, 2Department of Biomedical Engineering, The Johns Hopkins University, 3Department of Psychiatry, National Institute of Mental Health & Neuro Sciences (NIMHANS), 4Faculty of Computer Science, Dalhousie University, 5Department of Electrical and Computer Engineering, The Johns Hopkins University, 6Department of Computer Science, The Johns Hopkins University, 7Brain and Mind Institute, Department for Statistical and Actuarial Sciences, Department for Computer Science, Western University, 8Monash Biomedical Imaging, Monash University A standardized pipeline is presented for examining cerebellum grey matter morphometry. The pipeline combines high-resolution, state-of-the-art approaches for optimized and automated cerebellum parcellation and voxel-based registration of the cerebellum for volumetric quantification. Immunology and Infection Use of Viral Entry Assays and Molecular Docking Analysis for the Identification of Antiviral Candidates against Coxsackievirus A16 Jonathan Y. Wang1, Chien-Ju Lin2, Ching-Hsuan Liu3,4, Liang-Tzung Lin3,5 1Department of Molecular Biosciences, University of Texas at Austin, 2School of Pharmacy, College of Pharmacy, Kaohsiung Medical University, 3Graduate Institute of Medical Sciences, College of Medicine, Taipei Medical University, 4Department of Microbiology and Immunology, Dalhousie University, 5Department of Microbiology and Immunology, School of Medicine, College of Medicine, Taipei Medical University The goal of the protocol is to illustrate the different assays relating to viral entry that can be used to identify candidate viral entry inhibitors. Neuroscience Sagittal Plane Kinematic Gait Analysis in C57BL/6 Mice Subjected to MOG35-55 Induced Experimental Autoimmune Encephalomyelitis Maximillian DJ Fiander*1, Matthew AJ Chedrawe*1, Anna-Claire Lamport1, Turgay Akay2, George S Robertson1,3 1Pharmacology, Dalhousie University, 2Medical Neuroscience, Dalhousie University, 3Psychiatry, Dalhousie University Kinematic gait analysis in the sagittal plane yields highly precise information about how movement is executed. We describe the application of these techniques to identify gait deficits for mice subjected to autoimmune-mediated demyelination. These methods may also be used to characterize gait deficits for other mouse models featuring impaired locomotion. Neuroscience Immunohistochemical and Calcium Imaging Methods in Wholemount Rat Retina Allison Sargoy1, Steven Barnes1,2,3, Nicholas C. Brecha1,2,4, Luis Pérez De Sevilla Müller1 1Department of Neurobiology, University of California, Los Angeles, 2Veterans Administration Greater Los Angeles Healthcare System, 3Departments of Physiology & Biophysics and Ophthalmology & Visual Sciences, Dalhousie University, 4Departments of Neurobiology and Medicine, Jules Stein Eye Institute, CURE-Digestive Diseases Research Center, David Geffen School of Medicine, University of California, Los Angeles Immunohistochemistry protocols are used to study the localization of a specific protein in the retina. Calcium imaging techniques are employed to study calcium dynamics in retinal ganglion cells and their axons. Engineering Measurement of Coherence Decay in GaMnAs Using Femtosecond Four-wave Mixing Daniel Webber1, Tristan de Boer1, Murat Yildirim1, Sam March1, Reuble Mathew1, Angela Gamouras1, Xinyu Liu2, Margaret Dobrowolska2, Jacek Furdyna2, Kimberley Hall1 1Department of Physics and Atmospheric Science, Dalhousie University, 2Department of Physics, University of Notre Dame The technique of femtosecond four-wave mixing is described, including spectrally-resolved and time-resolved configurations. We illustrate the utility of this technique for the investigation of crucial physical properties in the III-V diluted magnetic semiconductors, afforded by its nonlinearity and high temporal resolution. Biology Horizontal Slice Preparation of the Retina Ryosuke Enoki1, Tatjana C. Jakobs2, Amane Koizumi2 1Dpt of Physiology and Biophysics, Dalhousie University, 2Massachusetts General Hospital, Harvard Medical School Traditionally the vertical slice and the whole-mount preparation of the retina have been used to study the function of retinal circuits. Here, we describe the novel slicing method to preserve the dendritic morphology of retinal neurons intact.