The Hospital for Sick Children 9 articles published in JoVE Immunology and Infection Scoring Central Nervous System Inflammation, Demyelination, and Axon Injury in Experimental Autoimmune Encephalomyelitis Carmen C. Ucciferri1, Annette Gower2, Nuria Alvarez-Sanchez1,2, Heather Whetstone3, Valeria Ramaglia1, Jennifer L. Gommerman1, Koroboshka Brand-Arzamendi2, Raphael Schneider2,4, Shannon E Dunn1,2,5 1Department of Immunology, University of Toronto, 2Keenan Research Centre for Biomedical Science of St. Michael’s Hospital, 3Sickkids Research Institute, The Hospital for Sick Children, 4Women’s College Research Institute, Women’s College Hospital Experimental autoimmune encephalomyelitis (EAE) serves as an animal model of multiple sclerosis. This article describes an approach for scoring spinal cord inflammation, demyelination, and axonal injury in EAE. Additionally, a method to quantify soluble neurofilament light levels in the mice serum is presented, facilitating the assessment of axonal injury in live mice. Cancer Research Magnetic Resonance-Guided High Intensity Focused Ultrasound Generated Hyperthermia: A Feasible Treatment Method in a Murine Rhabdomyosarcoma Model Claire Wunker1,2, Karolina Piorkowska3, Ben Keunen3, Yael Babichev2, Suzanne M. Wong3,4, Maximilian Regenold5, Michael Dunne5, Julia Nomikos1,2, Maryam Siddiqui6, Samuel Pichardo6, Warren Foltz7, Adam C. Waspe3,8, Justin T. Gerstle3,9, James M. Drake1,3,4,10, Rebecca A. Gladdy1,2,10 1Institute of Medical Science, University of Toronto, 2Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, 3The Wilfred and Joyce Posluns Centre for Image-Guided Innovation and Therapeutic Intervention, The Hospital for Sick Children, 4Institute of Biomedical Engineering, University of Toronto, 5Leslie Dan Faculty of Pharmacy, University of Toronto, 6Departments of Radiology and Clinical Neurosciences, University of Calgary, 7Department of Radiation Oncology, University of Toronto, 8Department of Medical Imaging, University of Toronto, 9Department of Pediatric Surgery, University of Toronto, 10Department of Surgery, University of Toronto Presented here is a protocol to use controlled hyperthermia, generated by magnetic resonance-guided high intensity focused ultrasound, to trigger drug release from temperature-sensitive liposomes in a rhabdomyosarcoma mouse model. Neuroscience Time-Lapse Imaging of Neuronal Arborization using Sparse Adeno-Associated Virus Labeling of Genetically Targeted Retinal Cell Populations Samantha Ing-Esteves1,2, Julie L. Lefebvre1,2 1Program for Neuroscience and Mental Health, Hospital for Sick Children, 2Department of Molecular Genetics, University of Toronto Here, we present a method for investigating neurite morphogenesis in postnatal mouse retinal explants by time-lapse confocal microscopy. We describe an approach for sparse labeling and acquisition of retinal cell types and their fine processes using recombinant adeno-associated virus vectors that express membrane-targeted fluorescent proteins in a Cre-dependent manner. Medicine Human Fetal Blood Flow Quantification with Magnetic Resonance Imaging and Motion Compensation Datta Singh Goolaub1,2, Davide Marini3,4, Mike Seed4,5, Christopher K. Macgowan1,2 1Department of Medical Biophysics, University of Toronto, 2Division of Translational Medicine, The Hospital for Sick Children, 3Labatt Family Heart Centre, The Hospital for Sick Children, 4Department of Pediatrics, University of Toronto, 5Division of Pediatric Cardiology, The Hospital for Sick Children Here we present a protocol for measuring fetal blood flow rapidly with MRI and retrospectively performing motion correction and cardiac gating. Biology Assessment of the Metabolic Effects of Isocaloric 2:1 Intermittent Fasting in Mice Ri Youn Kim*1,2, Ju Hee Lee*3,4, Yena Oh1,2, Hoon-Ki Sung3,4,5, Kyoung-Han Kim1,2 1University of Ottawa Heart Institute, 2Department of Cellular and Molecular Medicine, University of Ottawa, 3Translational Medicine Program, The Hospital for Sick Children, 4Department of Laboratory Medicine and Pathobiology, University of Toronto, 5Banting and Best Diabetes Centre, University of Toronto The current article describes a detailed protocol for isocaloric 2:1 intermittent fasting to protect and treat against obesity and impaired glucose metabolism in wild-type and ob/ob mice. Biology Visualization of 3D White Adipose Tissue Structure Using Whole-mount Staining Yanqing Jiang*1, Joanna Lan-Hing Yeung*1, Ju Hee Lee1,2, James An1, Patrick E. Steadman3, Jae-Ryong Kim4, Hoon-Ki Sung1,2,5 1Translational Medicine Program, The Hospital for Sick Children, 2Department of Laboratory Medicine and Pathobiology, University of Toronto, 3Neurosciences & Mental Health Program, The Hospital for Sick Children, 4Department of Biochemistry and Molecular Biology, Smart-Aging Convergence Research Center, College of Medicine, Yeungnam University, 5Banting and Best Diabetes Centre, University of Toronto The focus of the present study is to demonstrate the whole-mount immunostaining and visualization technique as an ideal method for 3D imaging of adipose tissue architecture and cellular component. Cancer Research Live-Cell Imaging Assays to Study Glioblastoma Brain Tumor Stem Cell Migration and Invasion Ian Restall1, Danielle Anne Bozek1, Charles Chesnelong1,2, Samuel Weiss1, H. Artee Luchman1 1Hotchkiss Brain Institute, Department of Cell Biology and Anatomy, Cumming School of Medicine, University of Calgary, 2Department of Developmental and Stem Cell Biology Program, Arthur and Sonia Labatt Brain Tumor Research Centre, The Hospital for Sick Children Here, we describe live-cell imaging techniques to quantitatively measure the migration and invasion of glioblastoma brain tumor stem cells over time and under multiple treatment conditions. Medicine Heterotopic Renal Autotransplantation in a Porcine Model: A Step-by-Step Protocol J. Moritz Kaths1,2, Juan Echeverri1,3, Nicolas Goldaracena1, Kristine S. Louis1, Paul Yip4, Rohan John4, Istvan Mucsi5, Anand Ghanekar1, Darius Bagli6, Markus Selzner1, Lisa A. Robinson2 1Multi Organ Transplant Program, Department of Surgery, Toronto General Hospital, 2Division of Nephrology, The Hospital for Sick Children, 3Programa de Doctorat en Medicina, La Universitat Autónoma de Barcelona, 4Laboratory Medicine and Pathobiology, Toronto General Hospital, 5Department of Medicine, Toronto General Hospital, 6Departments of Surgery (Urology) & Physiology, Developmental & Stem Cell Biology, The Hospital for Sick Children Porcine models of organ transplantation provide an important platform to study mechanisms of organ preservation. This article describes a heterotopic porcine renal autotransplantation model, which allows investigating new approaches to improve the outcome of transplantation using marginal kidney grafts. Developmental Biology Analysis of Zebrafish Larvae Skeletal Muscle Integrity with Evans Blue Dye Sarah J. Smith*1,2, Eric J. Horstick*3,4, Ann E. Davidson1,2, James Dowling1,2,4 1Program in Genetics & Genome Biology, The Hospital for Sick Children, 2Department of Molecular Genetics, The University of Toronto, 3Program in Genomics of Differentiation, Eunice Kennedy Shriver National Institute of Child Health and Human Development, 4Departments of Pediatrics and Neurology, University of Michigan In this study, we describe a straightforward method to perform Evans Blue Dye (EBD) analysis on zebrafish larvae. This technique is a powerful tool for the characterization of skeletal muscle integrity and delineation of zebrafish models of muscular dystrophy, and is a valuable method for the development of novel therapeutics.