University of Glasgow View Institution's Website 15 articles published in JoVE Biology Whole Genome Sequencing for Rapid Characterization of Rabies Virus Using Nanopore Technology Criselda Bautista1,2, Gurdeep Jaswant1,3,4,5, Hollie French1,6, Kathryn Campbell1, Rowan Durrant1, Robert Gifford1,6, Grace S. N. Kia7,8, Brian Ogoti3,9, Katie Hampson1, Kirstyn Brunker1,6 1School of Biodiversity, One Health & Veterinary Medicine, University of Glasgow, 2Research Institute for Tropical Medicine, 3University of Nairobi, Institute of Tropical and Infectious Diseases, 4Tanzania Industrial Research Development Organization, 5Ifakara Health Institute, 6MRC-University of Glasgow Centre for Virus Research, 7Department of Veterinary Public health, Ahmadu Bello University, 8African Centre of Excellence for Neglected Tropical Diseases and Forensic Biotechnology, Ahmadu Bello University, 9University of Nairobi, Kenya and Center for Epidemiological Modelling and Analysis, University of Nairobi Here, we present a rapid and cost-effective workflow for characterizing rabies virus (RABV) genomes using nanopore technology. The workflow is intended to support genomics-informed surveillance at a local level, providing information on circulating RABV lineages and their placement within regional phylogenies to guide rabies control measures. Neuroscience Establishing Mixed Neuronal and Glial Cell Cultures from Embryonic Mouse Brains to Study Infection and Innate Immunity Alistair Gamble1, Maria Suessmilch1, Anniek Bonestroo1, Andres Merits2, Gerard J. Graham1, Jonathan Cavanagh1, Julia M. Edgar1, Marieke Pingen1 1School of Infection and Immunity, University of Glasgow, 2Faculty of Science and Technology, Institute of Technology, University of Tartu This protocol presents a unique way of generating central nervous system cell cultures from embryonic day 17 mouse brains for neuro(immuno)logy research. This model can be analyzed using various experimental techniques, including RT-qPCR, microscopy, ELISA, and flow cytometry. Medicine Isolation and Culture of Human Adipose-Derived Stem Cells With an Innovative Xenogeneic-Free Method for Human Therapy Martino Guiotto*1,4, Silvia Palombella*2, Stefania Brambilla2, Lee Ann Applegate3, Mathis Riehle4, Andrew Hart4, Wassim Raffoul1, Pietro Giovanni di Summa1 1Department of Plastic, Reconstructive and Hand Surgery, Centre Hospitalier Universitaire Vaudois (CHUV, 2 Xenogeneic (chemical or animal-derived) products introduced in the cell therapy preparation/manipulation steps are associated with an increased risk of immune reactivity and pathogenic transmission in host patients. Here, a complete xenogeneic-free method for the isolation and in vitro expansion of human adipose-derived stem cells is described. Immunology and Infection Differentiation of Functional Osteoclasts from Human Peripheral Blood CD14+ Monocytes Patricia Riedlova*1, Shatakshi Sood*1, Carl S. Goodyear1, Cecilia Ansalone1 1School of Infection & Immunity, University of Glasgow Osteoclasts are key bone-resorbing cells in the body. This protocol describes a reliable method for the in vitro differentiation of osteoclasts from human peripheral blood monocytes. This method can be used as an important tool to further understand osteoclast biology in homeostasis and in diseases. Medicine Destabilization of the Medial Meniscus and Cartilage Scratch Murine Model of Accelerated Osteoarthritis Lynette Dunning1, Kendal McCulloch1, John C. Lockhart1, Carl S. Goodyear2, Carmen Huesa2 1Institute of Biomedical and Environmental Health Research, University of the West of Scotland, 2Institute of Infection, Immunity and Inflammation, University of Glasgow The present protocol describes the controlled microblade scratches on the surface of the articular cartilage after destabilizing the mouse knee by cutting the medial miniscotibial ligament. This animal model presents an accelerated form of osteoarthritis (OA) suitable for studying osteophyte formation, osteosclerosis, and early-stage pain. Medicine Interventional Diagnostic Procedure: A Practical Guide for the Assessment of Coronary Vascular Function Daniel T. Y. Ang1,2, Novalia P. Sidik1,2, Andrew J. Morrow1,2, Robert Sykes1,2, Margaret B. McEntegart1, Colin Berry1,2 1Department of Cardiology, Golden Jubilee National Hospital, 2British Heart Foundation Glasgow Cardiovascular Research Centre, University of Glasgow The purpose of this practical guide is to provide information on the preparation and administration of an interventional diagnostic procedure in clinical practice. It discusses some key preparation and safety considerations, as well as tips for procedural success. Bioengineering Experimental and Data Analysis Workflow for Soft Matter Nanoindentation Giuseppe Ciccone1, Mariana Azevedo Gonzalez Oliva1, Nelda Antonovaite2, Ines Lüchtefeld3, Manuel Salmeron-Sanchez1, Massimo Vassalli1 1Centre for the Cellular Microenvironment, James Watt School of Engineering, University of Glasgow, 2Optics 11 life, 3Laboratory of Biosensors and Bioelectronics, ETH Zürich The protocol presents a complete workflow for soft material nanoindentation experiments, including hydrogels and cells. First, the experimental steps to acquire force spectroscopy data are detailed; then, the analysis of such data is detailed through a newly developed open-source Python software, which is free to download from GitHub. Immunology and Infection A Tuberculosis Molecular Bacterial Load Assay (TB-MBLA) Wilber Sabiiti1, Bariki Mtafya1,2, Daniela Alferes De Lima1, Evelin Dombay1, Vincent O. Baron1, Khalide Azam3, Katarina Oravcova4, Derek J. Sloan1, Stephen H. Gillespie1 1Division of Infection and Global Health, School of Medicine, University of St Andrews, 2National Institute for Medical Research (NIMR)-Mbeya Medical Research Centre, 3Instituto Nacional de Saúde (INS), Ministério da Saúde, 4Institute of Biodiversity, Animal Health & Comparative Medicine, College of Medical, Veterinary & Life Sciences, University of Glasgow We describe a tuberculosis molecular bacterial load assay test performed after heat inactivation of sputum. Heat inactivation renders sputum samples noninfectious and obviates the need for containment level 3 laboratories for tuberculosis molecular tests. Cancer Research Subcellular Fractionation of Primary Chronic Lymphocytic Leukemia Cells to Monitor Nuclear/Cytoplasmic Protein Trafficking Jodie Hay*1, Michael W. Moles*1, Jennifer Cassels1, Alison M. Michie1 1Paul O'Gorman Leukaemia Research Centre, Institute of Cancer Sciences, College of Medical, Veterinary and Life Sciences, University of Glasgow This protocol enables the optimization and subsequent efficient generation of nuclear and cytoplasmic fractions from primary chronic lymphocytic leukemia cells. These samples are used to determine protein localization as well as changes in protein trafficking that take place between the nuclear and cytoplasmic compartments upon cell stimulation and drug treatment. Engineering Generation and Coherent Control of Pulsed Quantum Frequency Combs Benjamin MacLellan*1, Piotr Roztocki*1, Michael Kues1,2, Christian Reimer1, Luis Romero Cortés1, Yanbing Zhang1, Stefania Sciara1,3, Benjamin Wetzel1,4, Alfonso Cino3, Sai T. Chu5, Brent E. Little6, David J. Moss7, Lucia Caspani8, José Azaña1, Roberto Morandotti1,9,10 1Institut National de la Recherche Scientifique - Centre Énergie, Matériaux et Télécommunications (INRS-EMT), 2School of Engineering, University of Glasgow, 3Department of Energy, Information Engineering and Mathematical Models, University of Palermo, 4School of Mathematical and Physical Sciences, University of Sussex, 5Department of Physics and Material Science, City University of Hong Kong, 6State Key Laboratory of Transient Optics and Photonics, Xi'an Institute of Optics and Precision Mechanics, Chinese Academy of Science, 7Centre for Micro Photonics, Swinburne University of Technology, 8Institute of Photonics, Department of Physics, University of Strathclyde, 9Institute of Fundamental and Frontier Sciences, University of Electronic Science and Technology of China, 10National Research University of Information Technologies, Mechanics and Optics A protocol is presented for the practical generation and coherent manipulation of high-dimensional frequency-bin entangled photon states using integrated micro-cavities and standard telecommunications components, respectively. Medicine Performing Permanent Distal Middle Cerebral with Common Carotid Artery Occlusion in Aged Rats to Study Cortical Ischemia with Sustained Disability Christina Wayman*1,2, Denise A. Duricki*1,2, Lisa A. Roy3, Barbara Haenzi1, Shi-Yen Tsai4, Gwendolyn Kartje4,5,6, John S. Beech7, Diana Cash2, Lawrence Moon1 1Department of Neuroimaging, James Black Centre, Institute of Psychiatry, King's College London, University of London, 3Institute of Neuroscience and Psychology, Wellcome Surgical Institute, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, 4Research Service, Edward Hines Jr. VA Hospital, 5Neurology Service, Edward Hines Jr. VA Hospital, 6Department of Molecular Pharmacology and Therapeutics, Neuroscience Research Institute, Loyola University Chicago, 7Department of Oncology, The Gray Institute for Radiation, Oncology and Biology, University of Oxford Here we present a protocol to produce permanent distal middle cerebral artery occlusion in elderly female rats with simultaneous occlusion of the carotid arteries to generate large cortical infarcts and sustained deficits. We show confirmation of the lesion size using structural MRI at 24 hr and 8 weeks after stroke. Neuroscience Thermal Imaging to Study Stress Non-invasively in Unrestrained Birds Paul Jerem1, Katherine Herborn1, Dominic McCafferty1, Dorothy McKeegan1, Ruedi Nager1 1Institute of Biodiversity, Animal Health and Comparative Medicine, College of Medical, Veterinary & Life Sciences, University of Glasgow There is a need for a non-invasive assessment of stress. This paper describes a simple protocol using thermal imaging to detect a significant response in eye-region temperature in wild blue tits to a mild acute stressor. Bioengineering Sandwich-like Microenvironments to Harness Cell/Material Interactions José Ballester-Beltrán1,2, Myriam Lebourg1,3, Manuel Salmerón-Sánchez2 1Center for Biomaterials and Tissue Engineering, Universitat Politècnica de València, 2Division of Biomedical Engineering, School of Engineering, University of Glasgow, 3Biomedical Research Networking Center in Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN) The following protocol describes the procedure to assemble sandwich-like cultures to be used as an intermediate stage between bi-dimensional (2D) and three-dimensional (3D) cellular environments. The engineered systems can have applications in microscopy, biomechanics, biochemistry and cell biology assays. Engineering Simulation, Fabrication and Characterization of THz Metamaterial Absorbers James P. Grant1, Iain J.H. McCrindle1, David R.S. Cumming1 1School of Engineering, University of Glasgow This protocol outlines the simulation, fabrication and characterization of THz metamaterial absorbers. Such absorbers, when coupled with an appropriate sensor, have applications in THz imaging and spectroscopy. Bioengineering Organotypic Collagen I Assay: A Malleable Platform to Assess Cell Behaviour in a 3-Dimensional Context Paul Timpson1, Ewan J. Mcghee1, Zahra Erami1, Max Nobis1, Jean A. Quinn2, Mike Edward2, Kurt I. Anderson1 1The Beatson Institute for Cancer Research, University of Glasgow, 2Section of Dermatology, School of Medicine, University of Glasgow A method is described for the preparation of a 3-dimensional matrix consisting of collagen type I and primary human fibroblasts. This organotypic gel serves as a useful substrate to assess invasive cell migration because it mimics basic features of tissue stroma and is amenable to many forms of microscopy.
