University of London View Institution's Website 9 articles published in JoVE Biochemistry Preparing Lamellae from Vitreous Biological Samples Using a Dual-Beam Scanning Electron Microscope for Cryo-Electron Tomography Claudine Bisson1,2, Corey W. Hecksel3,4, James B. Gilchrist3, M. Alejandra Carbajal1, Roland A. Fleck1 1 Using focused ion beam milling to produce vitreous on-grid lamellae from plunge frozen biological samples for cryo-electron tomography. Medicine Cooling or Warming the Esophagus to Reduce Esophageal Injury During Left Atrial Ablation in the Treatment of Atrial Fibrillation Jason Zagrodzky1, Mark M. Gallagher2, Lisa W. M. Leung2, Tiffany Sharkoski3, Pasquale Santangeli4, Cory Tschabrunn4, Jose M. Guerra5, Bieito Campos5, John MacGregor6, Jamal Hayat2, Brad Clark7, Alex Mazur8, Marcel Feher9, Martin Arnold9, Mark Metzl10, Jose Nazari10, Erik Kulstad11 1 The goal of this protocol is to describe the use of esophageal temperature modulation to counteract esophageal thermal injury from left atrial ablation for the treatment of atrial fibrillation. Biology Measuring Lactase Enzymatic Activity in the Teaching Lab Cattleya S. Leksmono1, Claudia Manzoni1,2, James E. Tomkins1, Walter Lucchesi1,3, Graeme Cottrell1, Patrick A. Lewis1 1School of Pharmacy, University of Reading, 2Department of Molecular Neuroscience, UCL Institute of Neurology, 3School of Biological Sciences, Royal Hollway, University of London The enzymatic activity of lactase is essential for the catabolic processing of the disaccharide lactose. Here, the activity of lactase found in dietary supplements is assayed using a colorimetric assay. This provides students with an experimental platform for understanding the activity of lactase and enzyme kinetics. Developmental Biology Ex Vivo Culture of Chick Cerebellar Slices and Spatially Targeted Electroporation of Granule Cell Precursors Michalina Hanzel1, Richard J.T. Wingate1, Thomas Butts2 1 The cerebellar external granule layer is the site of the largest transit amplification in the developing brain. Here, we present a protocol to target genetic modification to this layer at the peak of proliferation using ex vivo electroporation and culture of cerebellar slices from embryonic Day 14 chick embryos. Environment Transcript and Metabolite Profiling for the Evaluation of Tobacco Tree and Poplar as Feedstock for the Bio-based Industry Colin Ruprecht1, Takayuki Tohge1, Alisdair Fernie1, Cara L. Mortimer2, Amanda Kozlo2, Paul D. Fraser2, Norma Funke1, Igor Cesarino3,4, Ruben Vanholme3,4, Wout Boerjan3,4, Kris Morreel3,4, Ingo Burgert5,6, Notburga Gierlinger5,6, Vincent Bulone7, Vera Schneider8, Andrea Stockero8, Juan Navarro-Aviñó9, Frank Pudel10, Bart Tambuyser11, James Hygate12, Jon Bumstead13, Louis Notley13, Staffan Persson1,14 1Max Planck Institute for Molecular Plant Physiology, 2School of Biological Sciences, Plant Molecular Science, Centre for Systems and Synthetic Biology, Royal Holloway, University of London, 3Department of Plant Systems Biology, VIB, 4Department of Plant Biotechnology and Bioinformatics, UGhent, 5Institute for Building Materials, ETH Zurich, 6Applied Wood Materials, EMPA, 7Division of Glycoscience, School of Biotechnology, AlbaNova University Center, Royal Institute of Technology (KTH), 8European Research and Project Office GmbH, 9ABBA Gaia S.L., 10Pflanzenöltechnologie, 11Capax Environmental Services, 12Green Fuels, 13Neutral Consulting Ltd, 14Plant Cell Biology Research Centre, School of Botany, University of Melbourne Plant biomass offers a renewable resource for multiple products, including fuel, feed, food, and a variety of materials. In this paper we investigate the properties of tobacco tree (Nicotiana glauca) and poplar as suitable sources for a biorefinery pipeline. Immunology and Infection A Functional Whole Blood Assay to Measure Viability of Mycobacteria, using Reporter-Gene Tagged BCG or M.Tb (BCG lux/M.Tb lux) Sandra Newton1, Adrian Martineau2, Beate Kampmann1 1Department of Paediatrics, Imperial College London, 2Centre for Health Sciences, Barts & The London School of Medicine and Dentistry We describe an alternative approach to the enumeration of mycobacteria in vitro, which uses reporter-gene tagged mycobacteria instead of colony-forming units (CFU). “Survival” of organisms as well as host response-markers are measured simultaneously, providing a low-cost, versatile and functional system for studies of host/pathogen interactions in the context of tuberculosis. Neuroscience Combining Behavioral Endocrinology and Experimental Economics: Testosterone and Social Decision Making Christoph Eisenegger*1, Michael Naef*2 1Institute for Empirical Research in Economics, Laboratory for Social and Neural Systems Research, University of Zurich, 2Department of Economics, Royal Holloway, University of London The procedure described in this protocol shows that testosterone administration and folk beliefs about testosterone may be associated with directly opposed social behaviors. Biology Studying Membrane Biogenesis with a Luciferase-Based Reporter Gene Assay Shaochong Zhang1, Axel Nohturfft1,2 1Department of Molecular and Cell Biology, Harvard, 2Molecular and Metabolic Signalling Centre, Division of Basic Medical Sciences, St. George's University of London Here, we describe procedures for studying changes in phagocytosis-induced gene expression with a luciferase-based reporter gene approach using the Dual-GloTM Luciferase Assay System from Promega. Biology Electroporation of Mycobacteria Renan Goude1, Tanya Parish1,2 1Center for Infectious Disease, Barts and the London School of Medicine and Dentistry, 2Institute for Cell and Molecular Science, Barts and the London School of Medicine and Dentistry Mycobacterial pathogenic strategies remain poorly understood. The slow growth rate of most species, the impenetrable nature of the cell-wall, and the hazards of working with pathogens make mycobacteria difficult to study and are largely responsible for our poor understanding of these organisms. In this video we will demonstrate the technique of electroporation, which involves subjecting cells to a brief high electrical impulse to allow the entry of DNA. It is the most widely used method for introducing DNA into mycobacterial cells.