University of Liverpool

16 articles published in JoVE

• Biology

  
  Determining Temperature Preference of Mosquitoes and Other Ectotherms Amirah Haziqah-Rashid¹, Kinga Stobierska¹, Lorna Glenn¹, Soeren Metelmann^1,2, Ken Sherlock³, Ewa Chrostek¹, Marcus S. C. Blagrove^{1 1}Department of Evolution, Ecology, and Behavior, Institute of Infection, Veterinary and Ecological Sciences, University of Liverpool, ²COVID-19 Outbreak Surveillance Team, UK Health Security Agency, ³Department of Livestock and One Health, Institute of Infection, Veterinary and Ecological Sciences, University of Liverpool Insects have an optimal environmental temperature range which they seek to remain within, and many external and internal factors can alter this preference. Here, we describe a cost-effective and simple method to study temperature choice, which allows insects to freely exhibit their natural behaviors.

• Chemistry

  
  Synthesis of Metal Nanoparticles Supported on Carbon Nanotube with Doped Co and N Atoms and its Catalytic Applications in Hydrogen Production Pui-Ching Poon*¹, Ka-Man Lee*¹, Yuanhao Wang², William Wai Yan Lam¹, Peter Shu-Wai Leung¹, Xiao-Ying Lu¹, Weiqun Li³, B. Layla Mehdi³, Yang Lu⁴, Chi-Wing Tsang¹, Eugene Yin Cheung Wong^{2 1}Faculty of Science and Technology, Technological and Higher Education Institute of Hong Kong (THEi), ²Department of Supply Chain and Information Management, The Hang Seng University of Hong Kong, ³Department of Mechanical, Materials and Aerospace Engineering, University of Liverpool, ⁴Laboratory of Building Environment and New Energy Resources, School of Civil Engineering, Faculty of Infrastructure Engineering, Dalian University of Technology Here, we present a protocol to synthesize Co nanoparticles supported on carbon nanotubes with Co- and N- dopants for hydrogen productions.

• Biochemistry

  
  An Economical and Versatile High-Throughput Protein Purification System Using a Multi-Column Plate Adapter Florence B Kineavy¹, Alex A Davies¹, Molly R Mitchell¹, Daniel Lay¹, Matt J Dominguez², Elliott J Stollar^{1 1}School of Life Sciences, University of Liverpool, ²Health Sciences Center, Texas Tech University A multi-column plate adapter allows chromatography columns to be interfaced with multi-well collection plates for parallel affinity or ion exchange purification providing an economical high throughput protein purification method. It can be used under gravity or vacuum yielding milligram quantities of protein via affordable instrumentation.

• Immunology and Infection

  
  Pan-lyssavirus Real Time RT-PCR for Rabies Diagnosis Denise A. Marston¹, Daisy L. Jennings¹, Nikki C. MacLaren¹, Daniel Dorey-Robinson¹, Anthony R. Fooks^1,2, Ashley C. Banyard¹, Lorraine M. McElhinney^{1,2 1}Wildlife Zoonoses & Vector-Borne Diseases Research Group, Animal and Plant Health Agency, ²Institute of Infection and Global Health, University of Liverpool This real-time RT-PCR using dsDNA intercalating dye is suitable to diagnose lyssavirus infections. The method begins with RNA extracted from rabies suspected ante-mortem or post-mortem samples, detailing master mix preparation, RNA addition, setup of the real-time machine and correct interpretation of results.

• Chemistry

  
  Reactive Inkjet Printing and Propulsion Analysis of Silk-based Self-propelled Micro-stirrers David A. Gregory¹, Piyush Kumar¹, Ana Jimenez-Franco¹, Yi Zhang¹, Yu Zhang ², Stephen J. Ebbens¹, Xiubo Zhao^{1 1}Department of Chemical and Biological Engineering, University of Sheffield, ²School of Engineering, University of Liverpool This protocol demonstrates the ability to utilize reactive inkjet printing to print self-motile biocompatible and environmentally friendly micro-stirrers for use in biomedical and environmental applications.

• Medicine

  
  Transdermal Measurement of Glomerular Filtration Rate in Mice Lauren Scarfe^1,2, Daniel Schock-Kusch³, Lorenzo Ressel⁴, Jochen Friedemann³, Yury Shulhevich³, Patricia Murray², Bettina Wilm², Mark de Caestecker^{1 1}Division of Nephrology, Department of Medicine, Vanderbilt University Medical Center, ²Department of Cellular and Molecular Physiology, University of Liverpool, ³MediBeacon GmbH, ⁴Department of Veterinary Pathology and Public Health, Institute of Veterinary Science, University of Liverpool Here we describe a protocol to measure glomerular filtration rate (GFR) in conscious, freely moving mice using a transdermal GFR monitor.

• Immunology and Infection

  
  Quantification of Efferocytosis by Single-cell Fluorescence Microscopy Kyle Taruc¹, Charles Yin¹, Daniel G. Wootton^2,3, Bryan Heit^{1 1}Department of Microbiology and Immunology and the Center for Human Immunology, University of Western Ontario, ²Institute of Infection and Global Health, University of Liverpool, ³Department of Respiratory Research, Aintree University Hospital NHS Foundation Trust Efferocytosis, the phagocytic removal of apoptotic cells, is required to maintain homeostasis and is facilitated by receptors and signaling pathways that allow for the recognition, engulfment, and internalization of apoptotic cells. Herein, we present a fluorescence microscopy protocol for the quantification of efferocytosis and the activity of efferocytic signaling pathways.

• Bioengineering

  
  Large-area Scanning Probe Nanolithography Facilitated by Automated Alignment and Its Application to Substrate Fabrication for Cell Culture Studies I-Ning Lee*^1,2, Joseph Hosford*¹, Shuai Wang³, John A. Hunt⁴, Judith M. Curran², William P. Heath³, Lu Shin Wong^{1 1}Manchester Institute of Biotechnology & School of Chemistry, University of Manchester, ²School of Engineering, University of Liverpool, ³School of Electrical and Electronic Engineering, University of Manchester, ⁴School of Science and Technology, Nottingham Trent University Here we present a protocol for wide-area scanning probe nanolithography enabled by the iterative alignment of probe arrays, as well as the utilization of lithographic patterns for cell-surface interaction studies.

• Medicine

  
  Contractility Measurements of Human Uterine Smooth Muscle to Aid Drug Development Sarah Arrowsmith*¹, Peter Keov², Markus Muttenthaler^3,4, Christian W. Gruber*^{2,5 1}Harris-Wellbeing Preterm Birth Research Centre, Department of Cellular and Molecular Physiology, Institute of Translational Medicine, University of Liverpool, ²School of Biomedical Sciences, The University of Queensland, ³Faculty of Chemistry, Institute of Biological Chemistry, University of Vienna, ⁴Institute for Molecular Bioscience, University of Queensland, ⁵Center for Physiology and Pharmacology, Medical University of Vienna This article describes experimental protocols to study ex vivo contractions of human myometrium and their application in drug discovery. This technique is used to improve the understanding of myometrial physiology and pathophysiology as well as to validate pharmacological data from novel research probes or drug leads.

• Developmental Biology

  
  Preparation and Culture of Myogenic Precursor Cells/Primary Myoblasts from Skeletal Muscle of Adult and Aged Humans Ana Soriano-Arroquia¹, Peter D. Clegg¹, Andrew P. Molloy^1,2, Katarzyna Goljanek-Whysall^{1 1}Institute of Ageing and Chronic Disease, University of Liverpool, ²Aintree University Hospital This protocol describes a robust, reproducible and simple method of isolation and culture of myoblast progenitor cells from the skeletal muscle of adult and aged people. The muscles used here include foot and leg muscles. This approach enables the isolation of an enriched population of primary myoblasts for functional studies.

• Biology

  
  Deferred Growth Inhibition Assay to Quantify the Effect of Bacteria-derived Antimicrobials on Competition Josephine C. Moran¹, Emma L. Crank¹, Hanaa A. Ghabban¹, Malcolm J. Horsburgh^{1 1}Institute of Integrative Biology, University of Liverpool The deferred growth inhibition assay can be used to assess the competition effect by one bacterial isolate on another. Inhibition is quantified by measuring the zone of clearing around the inhibitor-producing isolate, or qualitatively assessed by determining the visible extent of inhibition.

• Bioengineering

  
  Hollow Fiber Bioreactors for In Vivo-like Mammalian Tissue Culture Michael P. Storm¹, Ian Sorrell², Rebecca Shipley³, Sophie Regan², Kim A. Luetchford¹, Jean Sathish², Steven Webb⁴, Marianne J. Ellis^{1 1}Department of Chemical Engineering and Centre for Regenerative Medicine, University of Bath, ²MRC Centre for Drug Safety Science and Institute of Translational Medicine, University of Liverpool, ³Mechanical Engineering, University College London, ⁴Department of Applied Mathematics, Liverpool John Moores University The functional behavior of cells in culture can be improved by culturing in more in vivo-like 3-dimensional culture environments^16-21. This manuscript describes the set-up and operation of a hollow fiber bioreactor system for in vivo-like mammalian tissue culture.

• Neuroscience

  
  Dyeing Insects for Behavioral Assays: the Mating Behavior of Anesthetized Drosophila Rudi L. Verspoor¹, Chloe Heys¹, Thomas A. R. Price^{1 1}Institute of Integrative Biology, University of Liverpool This protocol describes a simple, cost effective way to individually identify Drosophila or other insects. Demonstration data investigating mating success across three species of Drosophila show that this method is comparable or better than the use of CO2 anaesthesia.

• Medicine

  
  Bronchoalveolar Lavage (BAL) for Research; Obtaining Adequate Sample Yield Andrea M. Collins^1,2, Jamie Rylance³, Daniel G. Wootton⁴, Angela D. Wright^3,5, Adam K. A. Wright^1,3, Duncan G. Fullerton^3,6, Stephen B. Gordon^{3 1}Biomedical Research Centre in Microbial Diseases, National Institute for Health Research, ²Respiratory Infection Group, Royal Liverpool and Broadgreen University Hospital Trust, ³Respiratory Infection Group, Liverpool School of Tropical Medicine, ⁴Institute of Infection and Global Health, University of Liverpool, ⁵Comprehensive Local Research Network, Royal Liverpool and Broadgreen University Hospital Trust, ⁶Department of Respiratory Research, University Hospital Aintree We describe a research technique for fiberoptic bronchoscopy and bronchoalveolar lavage using low pressure suction. The technique is used to harvest immune cells from the lung bronchoalveolar surfaces. Local anesthetic and mild conscious sedation (midazolam) is used. Subjects tolerate the procedure well and experience minimal side effects.

• Medicine

  
  Experimental Human Pneumococcal Carriage Jenna F. Gritzfeld¹, Angie D. Wright^1,2,3, Andrea M. Collins^1,2,4, Shaun H. Pennington¹, Adam K.A. Wright⁵, Aras Kadioglu⁶, Daniela M. Ferreira¹, Stephen B. Gordon^{1 1}Respiratory Infection Group, Liverpool School of Tropical Medicine, ²Royal Liverpool and Broadgreen, University Hospital Trust, ³Comprehensive Local Research Network, ⁴NIHR Biomedical Research Centre in Microbial Diseases, Royal Liverpool and Broadgreen University Hospitals NHS Trust, ⁵Institute of Lung Health, Respiratory Biomedical Unit, University Hospitals of Leicester NHS Trust & University of Leicester, ⁶Department of Clinical Infection Microbiology & Immunology, Institute of Infection & Global Health, University of Liverpool Experimental human pneumococcal carriage offers a natural model of carriage and a potential model for use in vaccine development. This technique is valuable yet complex and involves clinical risk by introducing a pathogen into a human. We have developed a detailed protocol.

• Immunology and Infection

  
  Use of Artificial Sputum Medium to Test Antibiotic Efficacy Against Pseudomonas aeruginosa in Conditions More Relevant to the Cystic Fibrosis Lung Sebastian Kirchner¹, Joanne L Fothergill², Elli A. Wright¹, Chloe E. James¹, Eilidh Mowat¹, Craig Winstanley^{1 1}Institute of Infection and Global Health, University of Liverpool, ²NIHR Biomedical Research Centre in Microbial Disease, University of Liverpool Current diagnostic antimicrobial susceptibility testing relies on the planktonic growth of isolates in nutrient rich, aerobic conditions. Here, we employ an alternative artificial sputum medium to study antimicrobial susceptibility of Pseudomonas aeruginosa biofilms under both aerobic and microaerophilic conditions more representative of the cystic fibrosis lung.