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University of Glasgow

16 ARTICLES PUBLISHED IN JoVE

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Bioengineering

Organotypic Collagen I Assay: A Malleable Platform to Assess Cell Behaviour in a 3-Dimensional Context
Paul Timpson 1, Ewan J. Mcghee 1, Zahra Erami 1, Max Nobis 1, Jean A. Quinn 2, Mike Edward 2, Kurt I. Anderson 1
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.

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JoVE Core

Simulation, Fabrication and Characterization of THz Metamaterial Absorbers
James P. Grant 1, Iain J.H. McCrindle 1, David R.S. Cumming 1
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.

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Bioengineering

Measurement of Tension Release During Laser Induced Axon Lesion to Evaluate Axonal Adhesion to the Substrate at Piconewton and Millisecond Resolution
Massimo Vassalli 1, Michele Basso 2, Francesco Difato 3
1Institute of Biophysics, National Research Council of Italy, 2Dipartimento di Sistemi e Informatica, Università di Firenze, 3Department of Neuroscience and Brain Technologies, Istituto Italiano di Tecnologia

We measured the tension release in an axon that was partially lesioned with a laser dissector by simultaneous force spectroscopy measurement performed on an optically-trapped probe adhered to the membrane of the axon. The developed experimental protocol evaluates the axon adhesion to the culture substrate.

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Bioengineering

Sandwich-like Microenvironments to Harness Cell/Material Interactions
José Ballester-Beltrán 1,2, Myriam Lebourg 1,3, Manuel Salmerón-Sánchez 2
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.

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Neuroscience

Thermal Imaging to Study Stress Non-invasively in Unrestrained Birds
Paul Jerem 1, Katherine Herborn 1, Dominic McCafferty 1, Dorothy McKeegan 1, Ruedi Nager 1
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.

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JoVE Core

Generation and Coherent Control of Pulsed Quantum Frequency Combs
Benjamin MacLellan *1, Piotr Roztocki *1, Michael Kues 1,2, Christian Reimer 1, Luis Romero Cortés 1, Yanbing Zhang 1, Stefania Sciara 1,3, Benjamin Wetzel 1,4, Alfonso Cino 3, Sai T. Chu 5, Brent E. Little 6, David J. Moss 7, Lucia Caspani 8, José Azaña 1, Roberto Morandotti 1,9,10
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.

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JoVE Journal

Subcellular Fractionation of Primary Chronic Lymphocytic Leukemia Cells to Monitor Nuclear/Cytoplasmic Protein Trafficking
Jodie Hay *1, Michael W. Moles *1, Jennifer Cassels 1, Alison M. Michie 1
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.

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Immunology and Infection

A Tuberculosis Molecular Bacterial Load Assay (TB-MBLA)
Wilber Sabiiti 1, Bariki Mtafya 1,2, Daniela Alferes De Lima 1, Evelin Dombay 1, Vincent O. Baron 1, Khalide Azam 3, Katarina Oravcova 4, Derek J. Sloan 1, Stephen H. Gillespie 1
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.

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Medicine

Interventional Diagnostic Procedure: A Practical Guide for the Assessment of Coronary Vascular Function
Daniel T. Y. Ang 1,2, Novalia P. Sidik 1,2, Andrew J. Morrow 1,2, Robert Sykes 1,2, Margaret B. McEntegart 1, Colin Berry 1,2
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.

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Bioengineering

Experimental and Data Analysis Workflow for Soft Matter Nanoindentation
Giuseppe Ciccone 1, Mariana Azevedo Gonzalez Oliva 1, Nelda Antonovaite 2, Ines Lüchtefeld 3, Manuel Salmeron-Sanchez 1, Massimo Vassalli 1
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.

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Medicine

Destabilization of the Medial Meniscus and Cartilage Scratch Murine Model of Accelerated Osteoarthritis
Lynette Dunning 1, Kendal McCulloch 1, John C. Lockhart 1, Carl S. Goodyear 2, Carmen Huesa 2
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.

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Immunology and Infection

Differentiation of Functional Osteoclasts from Human Peripheral Blood CD14+ Monocytes
Patricia Riedlova *1, Shatakshi Sood *1, Carl S. Goodyear 1, Cecilia Ansalone 1
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.

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Invasive Assessments For Coronary Vasomotor Disorders: Current State of the Art Methods Collection
Peter Ong 1, Colin Berry 2, Andreas Seitz 1
1Department of Cardiology and Angiology, Robert-Bosch-Krankenhaus, 2Department of Cardiology, Golden Jubilee National Hospital, British Heart Foundation Glasgow Cardiovascular Research Centre, University of Glasgow

Invasive Assessments For Coronary Vasomotor Disorders: Current State of the Art Methods Collection

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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 Brambilla 2, Lee Ann Applegate 3, Mathis Riehle 4, Andrew Hart 4, Wassim Raffoul 1, Pietro Giovanni di Summa 1
1Department of Plastic, Reconstructive and Hand Surgery, Centre Hospitalier Universitaire Vaudois (CHUV, 2Cell and Tissue Engineering Lab, IRCCS Ospedale Galeazzi Sant'Ambrogio, 3Unit of Regenerative Therapy, Plastic, Reconstructive and Hand Surgery, Department of Musculoskeletal Medicine, Lausanne University Hospital, 4Centre for the Cellular Microenvironment, University of Glasgow

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.

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Neuroscience

Establishing Mixed Neuronal and Glial Cell Cultures from Embryonic Mouse Brains to Study Infection and Innate Immunity
Alistair Gamble 1, Maria Suessmilch 1, Anniek Bonestroo 1, Andres Merits 2, Gerard J. Graham 1, Jonathan Cavanagh 1, Julia M. Edgar 1, Marieke Pingen 1
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.

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Biology

Whole Genome Sequencing for Rapid Characterization of Rabies Virus Using Nanopore Technology
Criselda Bautista 1,2, Gurdeep Jaswant 1,3,4,5, Hollie French 1,6, Kathryn Campbell 1, Rowan Durrant 1, Robert Gifford 1,6, Grace S. N. Kia 7,8, Brian Ogoti 3,9, Katie Hampson 1, Kirstyn Brunker 1,6
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.

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