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University of New South Wales

23 ARTICLES PUBLISHED IN JoVE

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Engineering

In Situ Neutron Powder Diffraction Using Custom-made Lithium-ion Batteries
William R. Brant 1, Siegbert Schmid 1, Guodong Du 2, Helen E. A. Brand 3, Wei Kong Pang 2,4,5, Vanessa K. Peterson 4, Zaiping Guo 2,5, Neeraj Sharma 6
1School of Chemistry, University of Sydney, 2Institute for Superconducting & Electronic Materials, University of Wollongong, 3Australian Synchrotron, 4Australian Nuclear Science and Technology Organisation, 5School of Mechanical, Materials, and Mechatronic Engineering, University of Wollongong, 6School of Chemistry, University of New South Wales

We describe the design and construction of an electrochemical cell for the examination of electrode materials using in situ neutron powder diffraction (NPD). We briefly comment on alternate in situ NPD cell designs and discuss methods for the analysis of the corresponding in situ NPD data produced using this cell.

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Bioengineering

Using Cell-substrate Impedance and Live Cell Imaging to Measure Real-time Changes in Cellular Adhesion and De-adhesion Induced by Matrix Modification
Martin D. Rees 1, Shane R. Thomas 1,2
1Centre for Vascular Research, University of New South Wales, 2School of Medical Sciences, University of New South Wales

Here, we present a protocol to continuously quantify cell adhesion and de-adhesion processes with high temporal resolution in a non-invasive manner by cell-substrate impedance and live cell imaging analyses. These approaches reveal the dynamics of cell adhesion/de-adhesion processes triggered by matrix modification and their temporal relationship to adhesion-dependent signaling events.

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Neuroscience

Intramuscular Injections Along the Motor End Plates: A Minimally Invasive Approach to Shuttle Tracers Directly into Motor Neurons
Rahul Mohan *1, Andrew P. Tosolini *1, Renée Morris 1
1Translational Neuroscience Facility, School of Medical Sciences, University of New South Wales

The efficacy of intramuscular uptake and retrograde transport of molecules to corresponding motor neurons depends on the location of the injection sites with respect to the motor end plates (MEPs). Here, we describe how to locate MEPs on skeletal muscles to optimise retrograde transport of tracers into motor neurons.

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JoVE Core

Silicon Metal-oxide-semiconductor Quantum Dots for Single-electron Pumping
Alessandro Rossi 1, Tuomo Tanttu 2, Fay E. Hudson 1, Yuxin Sun 1, Mikko Möttönen 2, Andrew S. Dzurak 1
1School of Electrical Engineering & Telecommunications, University of New South Wales, 2QCD Labs, COMP Centre of Excellence, Department of Applied Physics, Aalto University

The fabrication process and experimental characterization techniques relevant to single-electron pumps based on silicon metal-oxide-semiconductor quantum dots are discussed.

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Bioengineering

In Situ Mapping of the Mechanical Properties of Biofilms by Particle-tracking Microrheology
Su C. Chew 1,2, Scott A. Rice 2,3,4,5, Staffan Kjelleberg 2,3,4,6, Liang Yang 2,3
1Interdisciplinary Graduate School, Nanyang Technological University, 2Singapore Centre on Environmental Life Sciences Engineering, Nanyang Technological University, 3School of Biological Sciences, Nanyang Technological University, 4Centre for Marine Bio-Innovation, University of New South Wales, 5School of Biological, Earth and Environmental Sciences, University of New South Wales, 6School of Biotechnology and Biomolecular Sciences Sciences, University of New South Wales

Particle-tracking microrheology investigates the viscoelasticity of materials. Here, the technique is used to determine the viscoelasticity, creep compliance and effective crosslinking roles of different matrix components of a bacterial biofilm. The matrix consists of polymeric substances secreted by the bacteria and its components determine biofilm structure and mechanical properties.

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Chemistry

Facile Synthesis of Worm-like Micelles by Visible Light Mediated Dispersion Polymerization Using Photoredox Catalyst
Jonathan Yeow 1,2,3, Jiangtao Xu 1,2,3, Cyrille Boyer 1,2,3
1Centre for Advanced Macromolecular Design (CAMD), The University of New South Wales, 2Australian Centre for NanoMedicine (ACN), The University of New South Wales, 3School of Chemical Engineering, The University of New South Wales

This article describes a process for producing polymeric self-assembled nanoparticles using visible light mediated dispersion polymerization. Using low energy visible light to control the polymerization allows for the reproducible formation of self-assembled worm-like micelles at high solids content.

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Developmental Biology

Large-Scale Production of Cardiomyocytes from Human Pluripotent Stem Cells Using a Highly Reproducible Small Molecule-Based Differentiation Protocol
Hananeh Fonoudi *1,2,3,8, Hassan Ansari *1,8, Saeed Abbasalizadeh 1, Gillian M Blue 6,7, Nasser Aghdami 1, David S Winlaw 6,7, Richard P Harvey 2,3,4, Alexis Bosman *2,3, Hossein Baharvand *1,5
1Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, 2Developmental and Stem Cell Biology Division, Victor Chang Cardiac Research Institute, 3St. Vincent´s Clinical School, Faculty of Medicine, University of New South Wales, 4School of Biotechnology and Biomolecular Sciences, University of New South Wales, 5Department of Developmental Biology, University of Science and Culture, 6Heart Centre for Children, The Children´s Hospital at Westmead, 7Sydney Medical School, University of Sydney, 8Department of Developmental Biology, University of Science and Culture, Tehran, Iran

Here, we present a robust, fast and scalable cardiomyocyte differentiation protocol for human pluripotent stem cells (hPSCs). Cardiomyocytes derived using this large-scale method can provide sufficient cell numbers for their effective use in human cardiovascular disease modeling, high-throughput drug screening, and potentially clinical applications.

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Engineering

Recombination Dynamics in Thin-film Photovoltaic Materials via Time-resolved Microwave Conductivity
Joanna A. Guse 1,2, Timothy W. Jones 3, Andrew Danos 4, Dane R. McCamey 1,2
1ARC Centre of Excellence in Exciton Science, 2School of Physics, University of New South Wales, 3CSIRO, CSIRO Energy Centre, 4School of Chemistry, University of New South Wales

A Time Resolved Microwave Conductivity technique for investigating direct and trap-mediated recombination dynamics and determining carrier mobilities of thin film semiconductors is presented here.

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Developmental Biology

Generation of Genetically Modified Mice through the Microinjection of Oocytes
Fabien Delerue 1, Lars M. Ittner 1
1Transgenic Animal Unit, Mark Wainwright Analytical Centre, University of New South Wales

The microinjection of mouse oocytes is commonly used for both classic transgenesis (i.e., the random integration of transgenes) and CRISPR-mediated gene targeting. This protocol reviews the latest developments in microinjection, with a particular emphasis on quality control and genotyping strategies.

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Neuroscience

In Vivo Electrophysiological Measurement of the Rat Ulnar Nerve with Axonal Excitability Testing
Brandon M. Wild 1, Renée Morris 1, Mihai Moldovan 2, Christian Krarup 2, Arun V. Krishnan 3, Ria Arnold 1
1School of Medical Science, University of New South Wales, 2Department of Clinical Neurophysiology, Rigshospitalet and the Institute of Neuroscience and Pharmacology, University of Copenhagen, 3Prince of Wales Clinical School, University of New South Wales

Axonal excitability techniques provide a powerful tool to examine pathophysiology and biophysical changes that precede irreversible degenerative events. This manuscript demonstrates the use of these techniques on the ulnar nerve of anesthetized rats.

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Immunology and Infection

Megakaryocyte Differentiation and Platelet Formation from Human Cord Blood-derived CD34+ Cells
Jose Perdomo 1, Feng Yan 1, Halina H.L. Leung 1, Beng H. Chong 1,2
1Haematology Research Unit, St George and Sutherland Clinical School, University of New South Wales, 2Haematology Department, St George and Sutherland Hospitals

A highly pure population of megakaryocytes can be obtained from cord blood-derived CD34+ cells. A method for CD34+ cell isolation and megakaryocyte differentiation is described here.

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Immunology and Infection

Dissecting Multi-protein Signaling Complexes by Bimolecular Complementation Affinity Purification (BiCAP)
Jordan F. Hastings 1, Jeremy Z.R. Han 1, Robert F. Shearer 1,2, Sean P. Kennedy 1,3, Mary Iconomou 1,4, Darren N. Saunders 5, David R. Croucher 1,6,7
1The Kinghorn Cancer Centre, Garvan Institute of Medical Research, 2Ubiquitin Signaling Group, Protein Signaling Program, The Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, 3RCSI Molecular Medicine, Royal College of Surgeons in Ireland, 4Department of Epigenetics, Max Planck Institute of Immunobiology and Epigenetics, 5School of Medical Sciences, University of New South Wales, 6St Vincent's Hospital Clinical School, University of New South Wales, 7School of Medicine and Medical Science, University College Dublin

This manuscript describes the protocol for Bimolecular Complementation Affinity Purification (BiCAP). This novel method facilitates the specific isolation and downstream proteomic characterization of any two interacting proteins, while excluding un-complexed individual proteins as well as complexes formed with competing binding partners.

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Medicine

Laparoscopic Repair of Para-Esophageal Hernia Using Absorbable Biosynthetic Mesh
My Pham 1,2, Ruben Cohen-Hallaleh 1, Christophe R. Berney 1,2
1Department of General Surgery, Bankstown-Lidcombe Hospital, 2Medicine Faculty, University of New South Wales

Presented here is a protocol of para-esophageal hernia repair. Use of absorbable biosynthetic mesh avoids the risk of erosion through the esophagus whilst reinforcing the repair. Glue fixation is preferred to avoid the risk of trauma such as bleeding or cardiac tamponade, which are associated with stitches or tacks.

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Neuroscience

Combining Multiplex Fluorescence In Situ Hybridization with Fluorescent Immunohistochemistry on Fresh Frozen or Fixed Mouse Brain Sections
Ayse S. Dereli 1, Evan J. Bailey 1, Natasha N. Kumar 1
1Department of Pharmacology, School of Medical Sciences, University of New South Wales

This protocol describes a method for combining fluorescence in situ hybridization (FISH) and fluorescence immunohistochemistry (IHC) in both fresh frozen and fixed mouse brain sections, with the goal of achieving multilabel FISH and fluorescence IHC signal. IHC targeted cytoplasmic and membrane attached proteins.

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Cancer Research

An Orthotopic Resectional Mouse Model of Pancreatic Cancer
Tony C. Y. Pang 1,2,4,5, Zhihong Xu 1,2, Alpha Raj Mekapogu 1,2, Srinivasa Pothula 1,2, Therese M. Becker 3, David Goldstein 1, Romano C. Pirola 1,2, Jeremy S. Wilson 1,2, Minoti V. Apte 1,2
1Pancreatic Research Group, South Western Sydney Clinical School, University of New South Wales, 2Ingham Institute for Applied Medical Research, 3Centre for Circulating Tumour Cell Diagnostics and Research, Ingham Institute for Applied Medical Research, 4Surgical Innovations Unit, Westmead Hospital, 5Westmead Clinical School, University of Sydney

In the clinical context, patients with localized pancreatic cancer will undergo pancreatectomy followed by adjuvant treatment. This protocol reported here aims to establish a safe and effective method of modelling this clinical scenario in nude mice, through orthotopic implantation of pancreatic cancer followed by distal pancreatectomy and splenectomy.

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Neuroscience

Objectively Assessing Sports Concussion Utilizing Visual Evoked Potentials
Daryl H. Fong 1, Adrian J. Cohen 2, Dylan E. Mahony 2, Neil G. Simon 4, Joseph E. Herrera 3, Rebecca B. Baron 3, David Putrino 3
1School of Aerospace, Mechanical and Mechatronic Engineering, Faculty of Engineering and Information Technologies, University of Sydney, 2HeadsafeIP, 3Department of Rehabilitation Medicine, Icahn School of Medicine at Mount Sinai, 4St Vincent's Clinical School, Faculty of Medicine, University of New South Wales

A portable system capable of measuring steady-state visual-evoked potentials was developed and trialed on 65 amateur rugby players over 18 weeks to investigate SSVEP as a potential electrophysiological biomarker for concussion. Players' baselines were measured pre-season, with retesting for reliability, concussion, and recovery assessment being conducted within controlled time-periods, respectively.

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Medicine

Collection, Expansion, and Differentiation of Primary Human Nasal Epithelial Cell Models for Quantification of Cilia Beat Frequency
Katelin M. Allan 1,2, Sharon L. Wong 1,2, Laura K. Fawcett 1,2,3, Alexander Capraro 1,2, Adam Jaffe 1,2,3, Cristan Herbert 4, Elvis Pandzic 5, Shafagh A. Waters 1,2,3
1School of Women's and Children's Health, Faculty of Medicine and Health, University of New South Wales, 2Molecular and Integrative Cystic Fibrosis Research Centre (miCF RC), Faculty of Medicine and Health, University of New South Wales, 3Department of Respiratory Medicine, Sydney Children's Hospital, 4Department of Pathology, School of Medical Sciences, University of New South Wales Australia, 5Katharina Gaus Light Microscopy Facility, Mark Wainwright Analytical Centre, University of New South Wales

This protocol describes nasal epithelial cell collection, expansion, and differentiation to organotypic airway epithelial cell models and quantification of cilia beat frequency via live-cell imaging and custom-built scripts.

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Neuroscience

Expanding the Toolkit for In Vivo Imaging of Axonal Transport
Andrew P. Tosolini 1,2, David Villarroel-Campos 1,2,3, Giampietro Schiavo 1,2,3, James N. Sleigh 1,2,3
1Department of Neuromuscular Diseases, UCL Queen Square Institute of Neurology, University College London, 2UCL Queen Square Motor Neuron Disease Centre, University College London, 3UK Dementia Research Institute, University College London

Using transgenic fluorescent mice, detailed protocols are described to assess in vivo axonal transport of signaling endosomes and mitochondria within motor and sensory axons of the intact sciatic nerve in live animals.

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Chemistry

3D Printing and In Situ Surface Modification via Type I Photoinitiated Reversible Addition-Fragmentation Chain Transfer Polymerization
Nathaniel Corrigan 1, Cyrille Boyer 1
1Cluster for Advanced Macromolecular Design, and Australian Centre for Nanomedicine, School of Chemical Engineering, University of New South Wales

The present protocol describes the digital light processing-based 3D printing of polymeric materials using type I photoinitiated reversible addition-fragmentation chain transfer polymerization and the subsequent in situ material post-functionalization via surface-mediated polymerization. Photoinduced 3D printing provides materials with independently tailored and spatially controlled bulk and interfacial properties.

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Bioengineering

Ceramic Omnidirectional Bioprinting in Cell-Laden Suspensions for the Generation of Bone Analogs
Gagan Jalandhra 1, Sara Romanazzo 2, Stephanie Nemec 1, Iman Roohani 3, Kristopher A. Kilian 1,2
1School of Materials Science and Engineering, University of New South Wales, 2School of Chemistry, Australian Centre for NanoMedicine, University of New South Wales, 3School of Biomedical Engineering, University of Sydney

This protocol describes a 3D printing technique to fabricate bone-like structures by depositing a calcium phosphate ink in a gelatin-based granular support. Printed bone analogs are deposited in freeform, with flexibility for direct harvesting of the print or crosslinking within a living cell matrix for multiphasic constructs.

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Biology

A Semi-Automated Workflow for the Cryopreservation of Coral Sperm to Support Biobanking and Aquaculture
Jonathan Daly 1,2, Rebecca Hobbs 1, Nikolas Zuchowicz 3, Mary Hagedorn 4,5, Justine K. O’Brien 1,2
1Taronga Institute of Science and Learning, Taronga Conservation Society Australia, 2School of Biological, Earth and Environmental Sciences, University of New South Wales, 3Department of Mechanical Engineering, University of Minnesota, 4Hawaii Institute of Marine Biology, University of Hawaii, 5Smithsonian National Zoo and Conservation Biology Institute

This protocol describes a semi-automated pathway to improve the efficiency and capacity of processing and cryopreservation of sperm from threatened coral species, aiming to secure genetic diversity and support reef restoration efforts.

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Medicine

Using an Isolated Working Rat Heart Model to Test Donor Heart Preservation Strategies
Jeanette E. Villanueva 1,2, Ling Gao 1, Sanjay Dutta 1,2, Aoife Doyle 1, Yashutosh Joshi 1,2, Peter S. Macdonald 1,3,4
1Cardiac Physiology and Transplantation, Victor Chang Cardiac Research Institute, 2St Vincent's Healthcare Clinical Campus, University of New South Wales, 3Heart and Lung Transplant Unit, St Vincent's Hospital, 4Faculty of Medicine, University of New South Wales

We describe an ex vivo isolated working rat heart protocol to test donor heart preservation strategies. This paper describes the protocol for use in static cold storage of rodent donor hearts; however, the protocol can also be used for donor hearts obtained after donation after circulatory death and brain death.

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Biology

Long-term Monitoring of Oxygen Consumption Rates in Highly Differentiated and Polarized Retinal Pigment Epithelial Cultures
Qitao Zhang *1, Daisy Y. Shu *2, Richard A. Bryan 3, John Y. S. Han 1, Gillian A. Gulette 1, Kin Lo 3, Leo A. Kim 4, Jason M. L. Miller 1,5
1Kellogg Eye Center, University of Michigan, Ann Arbor, 2School of Optometry and Vision Science, University of New South Wales, 3Lucid Scientific, 4Schepens Eye Research Institute of Mass. Eye and Ear, Department of Ophthalmology, Harvard Medical School, 5Cellular and Molecular Biology Program, University of Michigan, Ann Arbor

We introduce a novel device for measuring oxygen consumption rates (OCR) in retinal pigment epithelial (RPE) cultures. The device can measure OCR for weeks at a time on RPE grown on standard cell culture plates with standard media while the plates are in a standard cell culture incubator.

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