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University of Sheffield

27 ARTICLES PUBLISHED IN JoVE

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Bioengineering

Induction of Adhesion-dependent Signals Using Low-intensity Ultrasound
James Roper 1, Andrew Harrison 2, Mark D. Bass 1
1School of Biochemistry, University of Bristol, 2Smith and Nephew

This protocol describes the stimulation of cultured fibroblasts with low-intensity pulsed ultrasound, which drives focal adhesion formation and Rac1 activation by mimicking engagement of the transmembrane matrix receptor, syndecan-4. This approach allows investigation of a successful clinical technique at the cellular level, thereby providing opportunities for refinement of the therapy.

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Bioengineering

Postproduction Processing of Electrospun Fibres for Tissue Engineering
Frazer J. Bye 1, Linge Wang 2, Anthony J. Bullock 1, Keith A. Blackwood 1, Anthony J. Ryan 3, Sheila MacNeil 1
1Materials Science and Engineering, University of Sheffield , 2Department of Biomedical Science, University of Sheffield , 3Department of Chemistry, University of Sheffield

Electrospun scaffolds can be processed post production for tissue engineering applications. Here we describe methods for spinning complex scaffolds (by consecutive spinning), for making thicker scaffolds (by multi-layering using heat or vapour annealing), for achieving sterility (aseptic production or sterilisation post production) and for achieving appropriate biomechanical properties.

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Engineering

Using Neutron Spin Echo Resolved Grazing Incidence Scattering to Investigate Organic Solar Cell Materials
Andrew J. Parnell 1, Adam Hobson 2, Robert M. Dalgliesh 3, Richard A. L. Jones 1, Alan D. F. Dunbar 2
1Department of Physics and Astronomy, University of Sheffield, 2Department of Chemical and Biological Engineering, The University of Sheffield, 3ISIS Pulsed Neutron and Muon Source Science and Technology Facilities Council, Rutherford Appleton Laboratory

Progress has been made in utilizing spin echo resolved grazing incidence scattering (SERGIS) as a neutron scattering technique to probe the length-scales in irregular samples. Crystallites of [6,6]-phenyl-C61-butyric acid methyl ester have been probed using the SERGIS technique and the results confirmed by optical and atomic force microscopy.

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Bioengineering

Combination of Microstereolithography and Electrospinning to Produce Membranes Equipped with Niches for Corneal Regeneration
Ílida Ortega 1, Farshid Sefat 1, Pallavi Deshpande 1, Thomas Paterson 1, Charanya Ramachandran 3, Anthony J. Ryan 2, Sheila MacNeil 1, Frederik Claeyssens 1
1Department of Materials Science and Engineering, University of Sheffield, 2Department of Chemistry, University of Sheffield, 3L. V. Prasad Eye Institute

We report a technique for the fabrication of micropockets within electrospun membranes in which to study cell behavior. Specifically, we describe a combination of microstereolithography and electrospinning for the production of PLGA (Poly(lactide-co-glycolide)) corneal biomaterial devices equipped with microfeatures.

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Bioengineering

Production, Characterization and Potential Uses of a 3D Tissue-engineered Human Esophageal Mucosal Model
Nicola H. Green 1, Bernard M. Corfe 2, Jonathan P. Bury 3, Sheila MacNeil 1
1Department of Materials Science and Engineering, University of Sheffield, 2Department of Oncology and Insigneo Institute for in silico Medicine, University of Sheffield, 3Department of Histopathology, Sheffield Teaching Hospitals NHS Foundation Trust

This manuscript describes the production, characterization and potential uses of a tissue engineered 3D esophageal construct prepared from normal primary human esophageal fibroblast and squamous epithelial cells seeded within a de-cellularized porcine scaffold. The results demonstrate the formation of a mature stratified epithelium similar to the normal human esophagus.

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Biology

Comparing the Affinity of GTPase-binding Proteins using Competition Assays
Rosalind C. Williamson 1, Mark D. Bass 2
1School of Biochemistry, University of Bristol, 2Department of Biomedical Science, University of Sheffield

This protocol compares the relative affinities of binding partners for Rho-family GTPases, including Rac1. In vivo, Rac1-binding proteins compete for a single binding interface, the conformation of which is dictated by a bound nucleotide. The nucleotide is both important and difficult to control experimentally, due to the high hydrolysis rate.

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Engineering

Blast Quantification Using Hopkinson Pressure Bars
Samuel D. Clarke 1, Stephen D. Fay 1,2, Samuel E. Rigby 1, Andrew Tyas 1,2, James A. Warren 1,2, Jonathan J. Reay 2, Benjamin J. Fuller 1, Matthew T. A. Gant 3, Ian D. Elgy 3
1Department of Civil & Structural Engineering, University of Sheffield, 2Blastech Ltd., 3Blast and IED Protection, Physical Sciences Department, Defence Science and Technology Laboratory (Dstl)

This protocol details the use of Hopkinson pressure bars to measure reflected blast loading from near-field explosive events. It is capable of interpolating a pressure-time history at any point on a reflective boundary and as such can be used to fully characterize the spatial and temporal variations in loading produced.

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JoVE Journal

Preparation and 3D Tracking of Catalytic Swimming Devices
Andrew Campbell 1, Richard Archer 1, Stephen Ebbens 1
1Department of Chemical and Biological Engineering, University of Sheffield

A method to prepare catalytically active Janus colloids that can "swim" in fluids and determine their 3D trajectories is presented.

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Chemistry

Laboratory Production of Biofuels and Biochemicals from a Rapeseed Oil through Catalytic Cracking Conversion
Siauw H. Ng 1, Yu Shi 1, Nicole E. Heshka 1, Yi Zhang 1, Edward Little 1
1CanmetENERGY, Natural Resources Canada

This paper presents an experimental method to produce biofuels and biochemicals from canola oil mixed with a fossil-based feed in the presence of a catalyst at mild temperatures. Gaseous, liquid, and solid products from a reaction unit are quantified and characterized. Conversion and individual product yields are calculated and reported.

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Neuroscience

In Vitro Recording of Mesenteric Afferent Nerve Activity in Mouse Jejunal and Colonic Segments
Sara Nullens 1, Annemie Deiteren 2, Wen Jiang 3, Christopher Keating 4, Hannah Ceuleers 1, Sven Francque 5, David Grundy 3, Joris G. De Man 1, Benedicte Y. De Winter 1
1Laboratory of Experimental Medicine and Pediatrics, Division of Gastroenterology, University of Antwerp, 2Visceral Pain Group, Discipline of Medicine, University of Adelaide, 3Department of Biomedical Sciences, University of Sheffield, 4Department of Pharmacy, Pharmacology and Postgraduate Medicine, University of Hertfordshire, 5Department of Gastroenterology and Hepatology, Antwerp University Hospital

Mesenteric afferent nerves convey information from the gastrointestinal tract towards the brain regarding normal homeostasis as well as pathophysiology. Gastrointestinal afferent nerve activity can be assessed by mounting isolated intestinal segments with attached afferent nerves into an organ bath, isolating the nerve, and assessing basal as well as stimulated activity.

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Bioengineering

Rapid Mix Preparation of Bioinspired Nanoscale Hydroxyapatite for Biomedical Applications
Caroline J. Wilcock 1, Piergiorgio Gentile 1, Paul V. Hatton 1, Cheryl A. Miller 1
1Bioengineering and Healthcare Technologies, School of Clinical Dentistry, University of Sheffield

This paper describes a novel method for the rapid manufacture of high quality bioinspired nanoscale hydroxyapatite. This biomaterial is of great significance in the manufacture of a wide range of innovative medical devices for clinical applications in orthopedics, craniofacial surgery and dentistry.

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Behavior

Use of a Video Scoring Anchor for Rapid Serial Assessment of Social Communication in Toddlers
Natasha Marrus 1, Stefanie Kennon-McGill 1, Brooke Harris 2, Yi Zhang 1, Anne L. Glowinski 1, John N. Constantino 1
1Department of Psychiatry, Washington University School of Medicine, 2Veteran's Administration Palo Alto Health Care System

Here we present a protocol for administration of the video-referenced rating of reciprocal social behavior (vrRSB) using a paper or online version. This survey quantifies RSB, a prerequisite for social competence, in toddlers through "video-referenced" items whereby a subject is compared to a reference child shown in a three-minute video.

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JoVE Core

Measuring the Carotid to Femoral Pulse Wave Velocity (Cf-PWV) to Evaluate Arterial Stiffness
Hongwei Ji 1, Jing Xiong 1, Shikai Yu 1, Chen Chi 1, Bin Bai 1, Jiadela Teliewubai 1, Yuyan Lu 1, Yi Zhang 1, Yawei Xu 1
1Department of Cardiology, Shanghai Tenth People's Hospital, Tongji University School of Medicine

This protocol describes a method to standardize the measurements of carotid-femoral pulse wave velocity to evaluate arterial stiffness.

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Chemistry

Preparation of Functional Silica Using a Bioinspired Method
Joseph R.H. Manning 1, Eleni Routoula 1, Siddharth V. Patwardhan 1
1Department of Chemical and Biological Engineering, University of Sheffield

Here, we present a protocol to synthesize bioinspired silica materials and immobilize enzymes therein. Silica is synthesized by combining sodium silicate and an amine 'additive', which neutralize at a controlled rate. Material properties and function can be altered either by in situ enzyme immobilization or post-synthetic acid elution of encapsulated additives.

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JoVE Core

Scalable Quantum Integrated Circuits on Superconducting Two-Dimensional Electron Gas Platform
Kaveh Delfanazari 1,2, Pengcheng Ma 2, Reuben Puddy 2, Teng Yi 2, Moda Cao 2, Yilmaz Gul 3, Carly L. Richardson 4, Ian Farrer 2,5, David Ritchie 2, Hannah J. Joyce 1, Michael J. Kelly 1,2, Charles G. Smith 2
1Centre for Advanced Photonics and Electronics, Engineering Department, University of Cambridge, 2Department of Physics, Cavendish Laboratory, University of Cambridge, 3Department of Electronic and Electrical Engineering, University College London, 4Department of Materials Science and Metallurgy, University of Cambridge, 5Department of Electronic and Electrical Engineering, University of Sheffield

Quantum integrated circuits (QICs) consisting of array of planar and ballistic Josephson junctions (JJs) based on In0.75Ga0.25As two-dimensional electron gas (2DEG) is demonstrated. Two different methods for fabrication of the two-dimensional (2D) JJs and QICs are discussed followed by the demonstration of quantum transport measurements in sub-Kelvin temperatures.

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Cancer Research

Orthotopic Injection of Breast Cancer Cells into the Mice Mammary Fat Pad
Gan-Lin Zhang 1, Yi Zhang 1, Ke-Xin Cao 1, Xiao-Min Wang 1
1Oncology Department, Beijing Hospital of Traditional Chinese Medicine, Capital Medical University

Here, we present a protocol to implant breast cancer cells into the mammary fat pad in a simple, less invasive, and easy-to-handle way, and this mouse orthotopic breast cancer model with a proper mammary fat pad environment can be used to investigate various aspects of cancer.

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Chemistry

Reactive Inkjet Printing and Propulsion Analysis of Silk-based Self-propelled Micro-stirrers
David A. Gregory 1, Piyush Kumar 1, Ana Jimenez-Franco 1, Yi Zhang 1, Yu Zhang  2, Stephen J. Ebbens 1, Xiubo Zhao 1
1Department 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.

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Chemistry

A Femtoliter Droplet Array for Massively Parallel Protein Synthesis from Single DNA Molecules
Yi Zhang 1, Kanako Kurosawa 1, Daisuke Nishiura 2, Mika Tei 1, Mikiko Tsudome 3
1SUGAR Program, X-star, Japan Agency for Marine-Earth Science and Technology (JAMSTEC), 2Center for Mathematical Science and Advanced Technology, Japan Agency for Marine-Earth Science and Technology (JAMSTEC), 3Deep-Sea Nanoscience Research Group, Japan Agency for Marine-Earth Science and Technology (JAMSTEC)

The overall goal of the protocol is to prepare over one million ordered, uniform, stable, and biocompatible femtoliter droplets on a 1 cm2 planar substrate that can be used for cell-free protein synthesis.

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Immunology and Infection

Establishing a Porcine Ex Vivo Cornea Model for Studying Drug Treatments against Bacterial Keratitis
Katarzyna Okurowska 1,2, Sanhita Roy 3, Praveen Thokala 4, Lynda Partridge 1,5, Prashant Garg 3, Sheila MacNeil 1,6, Peter N. Monk 1,7, Esther Karunakaran 1,2
1Sheffield Collaboratorium for Antimicrobial Resistance and Biofilms (SCARAB), University of Sheffield, 2Department of Chemical and Biological Engineering, University of Sheffield, 3Hyderabad Eye Research Foundation, L V Prasad Eye Institute, 4Health Economics and Decision Science, School of Health and Related Research, University of Sheffield, 5Department of Molecular Biology and Biotechnology, University of Sheffield, 6Department of Materials Science and Engineering, University of Sheffield, 7Department of Infection, Immunity and Cardiovascular Disease, University of Sheffield

This article describes a step-by-step protocol to set up an ex vivo porcine model of bacterial keratitis. Pseudomonas aeruginosa is used as a prototypic organism. This innovative model mimics in vivo infection as bacterial proliferation is dependent on the ability of the bacterium to damage corneal tissue.

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Biochemistry

Making Precise and Accurate Single-Molecule FRET Measurements using the Open-Source smfBox
Mahmoud A. S. Abdelhamid *1, Alice V. Rhind-Tutt *1, Benjamin Ambrose 1, Timothy D. Craggs 1
1Sheffield Institute for Nucleic Acids, Department of Chemistry, University of Sheffield

This article provides step-by-step instructions for making fully-corrected accurate FRET measurements on individual, freely diffusing biomolecules using the open-source, inexpensive smfBox, from switch on, through alignment and focusing, to data collection and analysis.

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Biochemistry

Magnetic Tweezers in a Microplate Format
Ália dos Santos 1, Christopher P. Toseland 1
1Department of Oncology and Metabolism, University of Sheffield

Here, we describe the use of a novel microplate assay to enable mechanical manipulation of biomolecules while performing ensemble biochemical assays. This is achieved using a microplate lid modified with magnets to create multiple static magnetic tweezers across the microplate.

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Immunology and Infection

A Novel High-Throughput Ex Vivo Ovine Skin Wound Model for Testing Emerging Antibiotics
Hannah C. Regan 1,2, Annette F. Taylor 1,2, Esther Karunakaran 1,2
1Sheffield Collaboratorium for Antimicrobial Resistance and Biofilms (SCARAB), University of Sheffield, 2Department of Chemical and Biological Engineering, University of Sheffield

The protocol describes a step-by-step method to set up an ex vivo ovine wounded skin model infected with Staphylococcus aureus. This high-throughput model better simulates infections in vivo compared with conventional microbiology techniques and presents researchers with a physiologically relevant platform to test the efficacy of emerging antimicrobials.

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Neuroscience

Real-Time Dynamic Collection of Hippocampal Extracellular Fluid from Conscious Rats Using a Microdialysis System
Xiaobo Wang 1,2, Na Xie 3, Yi Zhang 4, Xianli Meng 1,2,3, Ya Hou 3, Sanyin Zhang 1,2
1Innovative Institute of Chinese Medicine and Pharmacy, Chengdu University of Traditional Chinese Medicine, 2Research Institute of Integrated TCM & Western Medicine, Chengdu University of Traditional Chinese Medicine, 3School of Pharmacy, Chengdu University of Traditional Chinese Medicine, 4School of Ethnic Medicine, Chengdu University of Traditional Chinese Medicine

The protocol here provides a detailed real-time dynamic sampling of extracellular fluid from the hippocampus of awake rats using a microdialysis system.

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Medicine

Dynamic Continuous Blood Extraction from Rat Heart via Noninvasive Microdialysis Technique
Ya Hou 1, Jinrong Bai 1,2, Yi Zhang 3, Xianli Meng 1,2,4, Sanyin Zhang 4, Xiaobo Wang 2
1School of Pharmacy, Chengdu University of Traditional Chinese Medicine, 2Research Institute of Integrated TCM & Western Medicine, Chengdu University of Traditional Chinese Medicine, 3School of Ethnic Medicine, Chengdu University of Traditional Chinese Medicine, 4Innovative Institute of Chinese Medicine and Pharmacy, Chengdu University of Traditional Chinese Medicine

The present protocol describes a simple and efficient method for the real-time and dynamic collection of rat heart blood using the microdialysis technique.

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JoVE Core

An In Vitro Dissolution Determination of Multi-Index Components in Tibetan Medicine Rhodiola Granules
Qinyun Du 1, Qingxiu He 2, Fengming Zhang 3, Jiao Mi 3, Yuying Li 3, Shaohui Wang 1, Yi Zhang 1
1School of Pharmacy, Chengdu University of Traditional Chinese Medicine, 2School of Ethnic Medicine, Chengdu University of Traditional Chinese Medicine, 3Tibet Rhodiola Pharmaceutical Holding Co. Ltd

Here, we test the dissolution of Rhodiola granules (RG) in vitro, draw dissolution curves of salidroside, gallic acid, and ethyl gallate in ultrapure water, and fit the curves to different mathematical models. This protocol provides information and guidance for in vivo bioequivalence and in vivo-in vitro correlation studies of RG.

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Bioengineering

Synthesis of Strong Adhesive Hydrogel, Gelatin O-Nitrosobenzaldehyde
Yuelong Liang *1,2, Zhengze Huang *1, Yi Zhang *3,4, Yi Hong 3,4, Qijiang Mao 1,2,5, Xu Feng 1
1Department of General Surgery, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, 2Zhejiang Province Medical Research Center of Minimally Invasive Diagnosis and Treatment of Abdominal Diseases, 3Dr. Li Dak Sum & Yip Yio Chin Center for Stem Cells and Regenerative Medicine, Department of Orthopedic Surgery of the Second Affiliated Hospital, Zhejiang University School of Medicine, 4Zhejiang University-University of Edinburgh Institute, Zhejiang University School of Medicine, Key Laboratory of Tissue Engineering and Regenerative Medicine of Zhejiang Province, Zhejiang University School of Medicine, 5Zhejiang Provincial Key Laboratory of Laparoscopic Technology

The protocol presented here shows the synthesis of a strong adhesive hydrogel gelatin o-nitrosobenzaldehyde (gelatin-NB). Gelatin-NB has rapid and efficient tissue adhesion ability, which can form a strong physical barrier to protect wound surfaces, so it is expected to be applied to the field of injury repair biotechnology.

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Medicine

Voltage-Dependent Potassium Current Recording on H9c2 Cardiomyocytes via the Whole-Cell Patch-Clamp Technique
Hong Jiang *1, Yating Zhang *1, Ya Hou 1, Lijun Li 2, Sanyin Zhang 3,4, Yi Zhang 5, Xianli Meng 1,3,4, Xiaobo Wang 4
1State Key Laboratory of Southwestern Chinese Medicine Resources, School of Pharmacy, Chengdu University of Traditional Chinese Medicine, 2Chengdu Techman Instrument Co., Ltd., 3Innovative Institute of Chinese Medicine and Pharmacy, Chengdu University of Traditional Chinese Medicine, 4Research Institute of Integrated TCM & Western Medicine, Chengdu University of Traditional Chinese Medicine, 5School of Ethnic Medicine, Chengdu University of Traditional Chinese Medicine

The present protocol describes an efficient method for the real-time and dynamic acquisition of voltage-gated potassium (Kv) channel currents in H9c2 cardiomyocytes using the whole-cell patch-clamp technique.

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