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La Trobe University

19 ARTICLES PUBLISHED IN JoVE

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JoVE Core

Behavioral Determination of Stimulus Pair Discrimination of Auditory Acoustic and Electrical Stimuli Using a Classical Conditioning and Heart-rate Approach
Simeon J. Morgan 1, Antonio G. Paolini 1
1School of Psychological Science, La Trobe University

The application of a classical fear conditioning behavioral paradigm for auditory prosthetic research in rats is described. This paradigm provides a mechanism for identifying both detection of, and discrimination between, distinct acoustic and electrical stimuli using heart-rate as an outcome measure.

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Neuroscience

Thinned-skull Cortical Window Technique for In Vivo Optical Coherence Tomography Imaging
Jenny I. Szu 1, Melissa M. Eberle 2, Carissa L. Reynolds 2, Mike S. Hsu 1, Yan Wang 2, Christian M. Oh 2, M. Shahidul Islam 2, B. Hyle Park 2, Devin K. Binder 1
1Division of Biomedical Sciences, University of California, Riverside , 2Department of Bioengineering, University of California, Riverside

We present a method of creating a thinned-skull cortical window (TSCW) in a mouse model for in vivo OCT imaging of the cerebral cortex.

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Neuroscience

A Method for Systematic Electrochemical and Electrophysiological Evaluation of Neural Recording Electrodes
Alexander R. Harris 1,3, Simeon J. Morgan 1,3, Gordon G. Wallace 3, Antonio G. Paolini 1,3,4
1School of Psychological Science, La Trobe University, 2Intelligent Polymer Research Institute, University of Wollongong, 3ARC Centre of Excellence for Electromaterials Science, 4Health Innovations Research Institute, College of Science, Engineering, and Health, RMIT University

Different electrode coatings affect neural recording performance through changes to electrochemical, chemical and mechanical properties. Comparison of electrodes in vitro is relatively simple, however comparison of in vivo response is typically complicated by variations in electrode/neuron distance and between animals. This article provides a robust method to compare neural recording electrodes.

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Medicine

Rat Model of Photochemically-Induced Posterior Ischemic Optic Neuropathy
Yan Wang 1,3,4, Dale P. Brown 1, Brant D. Watson 2, Jeffrey L. Goldberg 1,3
1Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, 2Departments of Neurology and Biomedical Engineering, University of Miami Miller School of Medicine, 3Shiley Eye Center, University of California, 4Department of Ophthalmology and Vision Science, Fudan University

The goal of this protocol is to photochemically induce ischemic injury to the posterior optic nerve in rat. This model is critical to studies of the pathophysiology of posterior ischemic optic neuropathy, and therapeutic approaches for this and other optic neuropathies, as well as of other CNS ischemic diseases.

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Environment

Functionalization and Dispersion of Carbon Nanomaterials Using an Environmentally Friendly Ultrasonicated Ozonolysis Process
Eudora S. Y. Yeo 1, Gary I. Mathys 2, Narelle Brack 3, Erik T. Thostenson 4,5,6, Andrew N. Rider 1
1Aerospace Division, Defence Science and Technology Group, 2Maritime Division, Defence Science and Technology Group, 3Department of Chemistry and Physics, La Trobe Institute for Molecular Science, La Trobe University, 4Department of Mechanical Engineering, University of Delaware, 5Department of Materials Science and Engineering, University of Delaware, 6Center for Composite Materials, University of Delaware

Here, a novel method for the functionalization and stable dispersion of carbon nanomaterials in aqueous environments is described. Ozone is injected directly into an aqueous dispersion of carbon nanomaterial that is continuously recirculated through a high-powered ultrasonic cell.

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Chemistry

Measurements of Long-range Electronic Correlations During Femtosecond Diffraction Experiments Performed on Nanocrystals of Buckminsterfullerene
Rebecca A. Ryan 1, Sophie Williams 1, Andrew V. Martin 1, Ruben A. Dilanian 1, Connie Darmanin 2, Corey T. Putkunz 1, David Wood 3, Victor A. Streltsov 4, Michael W.M. Jones 5, Naylyn Gaffney 6, Felix Hofmann 7, Garth J. Williams 8, Sebastien Boutet 9, Marc Messerschmidt 10, M. Marvin Seibert 11, Evan K. Curwood 11, Eugeniu Balaur 2, Andrew G. Peele 5, Keith A. Nugent 2, Harry M. Quiney 1, Brian Abbey 2
1ARC Centre of Excellence in Advanced Molecular Imaging, School of Physics, University of Melbourne, 2Australian Research Council (ARC) Centre of Excellence in Advanced Molecular Imaging, Department of Chemistry and Physics, La Trobe Institute for Molecular Sciences, La Trobe University, 3Department of Physics, Imperial College London, 4Florey Institute of Neuroscience and Mental Health, 5Science and Engineering Faculty, Queensland University of Technology, 6Swinburne University of Technology, 7Department of Engineering Science, University of Oxford, 8Brookhaven National Laboratory, 9Linac Coherent Light Source, SLAC National Accelerator Laboratory, 10BioXFEL Science and Technology Center, 11Laboratory of Molecular Biophysics, Department of Cell and Molecular Biology, Uppsala University, 12Australian Synchrotron

We describe an experiment designed to probe the electronic damage induced in nanocrystals of Buckminsterfullerene (C60) by intense, femtosecond pulses of X-rays. The experiment found that, surprisingly, rather than being stochastic, the X-ray induced electron dynamics in C60 are highly correlated, extending over hundreds of unit cells within the crystals1.

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Bioengineering

Development of an Electrochemical DNA Biosensor to Detect a Foodborne Pathogen
Noordiana Nordin 1, Nor Azah Yusof 2,3, Son Radu 1, Roozbeh Hushiarian 4
1Laboratory of Food Safety and Food Integrity, Institute of Tropical Agriculture and Food Security, Universiti Putra Malaysia, 2Laboratory of Functional Device, Institute of Advanced Technology, Universiti Putra Malaysia, 3Department of Chemistry, Faculty of Science, Universiti Putra Malaysia, 4La Trobe Institute for Molecular Science, La Trobe University

A protocol for the development of an electrochemical DNA biosensor comprising a polylactic acid-stabilized, gold nanoparticles-modified, screen-printed carbon electrode to detect Vibrio parahaemolyticus is presented.

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JoVE Journal

Isolation and Respiratory Measurements of Mitochondria from Arabidopsis thaliana
Wenhui Lyu 1, Jennifer Selinski 1, Lu Li 1, David A. Day 2, Monika W. Murcha 3, James Whelan 1, Yan Wang 1
1ARC Centre of Excellence in Plant Energy Biology, Department of Animal, Plant and Soil Science, School of Life Science, La Trobe University, 2School of Biological Sciences, Flinders University, 3ARC Centre of Excellence in Plant Energy Biology, University of Western Australia

As mitochondria are only a small percentage of the plant cell, they need to be purified for a range of studies. Mitochondria can be isolated from a variety of plant organs by homogenization, followed by differential and density gradient centrifugation to obtain a highly purified mitochondrial fraction.

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Biology

Confocal Microscopy Reveals Cell Surface Receptor Aggregation Through Image Correlation Spectroscopy
Adam C. Parslow 1,2, Andrew H.A. Clayton 3, Peter Lock 4, Andrew M. Scott 1,2,5,6,7
1Tumour Targeting Laboratory, Olivia Newton-John Cancer Research Institute, 2School of Cancer Medicine, La Trobe University, 3Centre for Micro-Photonics, Faculty of Science, Engineering and Technology, Swinburne University of Technology, 4LIMS Bioimaging Facility, La Trobe Institute for Molecular Science, La Trobe University, 5Department of Medical Oncology, Olivia Newton-John Cancer and Wellnes Centre, Austin Health, 6Department of Medicine, University of Melbourne, 7Department of Molecular Imaging and Therapy, Austin Health

Antibodies that bind to target receptors on the cell surface can confer conformation and clustering alterations. These dynamic changes have implications for characterizing drug development in target cells. This protocol utilizes confocal microscopy and image correlation spectroscopy through ImageJ/FIJI to quantify the extent of receptor clustering on the cell surface.

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Biochemistry

Detection and Isolation of Apoptotic Bodies to High Purity
Thanh Kha Phan 1, Ivan KH Poon 1, Georgia K Atkin-Smith 1
1Department of Biochemistry and Genetics, La Trobe Institute for Molecular Science, La Trobe University

A workflow using flow cytometry or differential centrifugation is developed to detect, quantify and isolate apoptotic bodies from an apoptotic sample to high purity.

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Medicine

Non-invasive Assessment of Dorsiflexor Muscle Function in Mice
Frederico Gerlinger-Romero 1, Alex B. Addinsall 2, Richard M. Lovering 3, Victoria C. Foletta 4, Chris van der Poel 5, Paul A. Della-Gatta 4, Aaron P. Russell 4
1School of Exercise and Nutrition Sciences, Deakin University, 2Centre for Molecular and Medical Research, School of Medicine, Deakin University, 3Department of Orthopaedics, School of Medicine, University of Maryland, 4Institute for Physical Activity and Nutrition (IPAN), School of Exercise and Nutrition Sciences, Deakin University, 5Department of Physiology, Anatomy and Microbiology, La Trobe University

Measurement of rodent skeletal muscle contractile function is a useful tool that can be used to track disease progression as well as efficacy of therapeutic intervention. We describe here the non-invasive, in vivo assessment of the dorsiflexor muscles that can be repeated over time in the same mouse.

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Medicine

Protocol and Guidelines for Point-of-Care Lung Ultrasound in Diagnosing Neonatal Pulmonary Diseases Based on International Expert Consensus
Jing Liu 1,2, Roberto Copetti 3, Erich Sorantin 4, Jovan Lovrenski 5, Javier Rodriguez-Fanjul 6, Dalibor Kurepa 7, Xing Feng 8, Luigi Cattaross 9, Huayan Zhang 10,11, Misun Hwang 12, Tsu F. Yeh 13,14, Yisrael Lipener 7, Abhay Lodha 15, Jia-Qin Wang 16, Hai-Ying Cao 2,17, Cai-Bao Hu 2,18, Guo-Rong Lyu 19, Xin-Ru Qiu 1,2, Li-Qun Jia 20, Xiao-Man Wang 20, Xiao-Ling Ren 1,2, Jiu-Ye Guo 1,2, Yue-Qiao Gao 1,2, Jian-Jun Li 1,2, Ying Liu 1,2, Wei Fu 1,2, Yan Wang 21, Zu-Lin Lu 1,2, Hua-Wei Wang 8, Li-Li Shang 22
1Department of Neonatology and NICU, Beijing Chaoyang District Maternal and Child Healthcare Hospital, 2The Neonatal Lung Ultrasound Training Base, Chinese College of Critical Ultrasound, 3Emergency Department, Cattinara University Hospital, 4Division of Pediatric Radiology, Department of Radiology, Medical University Graz, 5Faculty of Medicine, University of Novi Sad, Radiology Department, Institute for Children and Adolescents Health Care of Vojvodina, 6Pediatric Intensive Care Unit, Pediatric Service Hospital Joan XXIII Tarragona, University Rovira i Virgil, 7Division of Neonatal-Perinatal Medicine, Cohen Children's Medical Center, 8Department of Neonatology, Children's Hospital of Soochow University, 9Department of Neonatology, Udine University Hospital, 10Center for Newborn Care, Guangzhou Women and Children's Medical Center, 11Division of Neonatology, Children's Hospital of Philadelphia, 12Department of Radiology, Children's Hospital of Philadelphia, 13Division of Neonatology and NICU, Cook County Children's Hospital, University of Illinois, 14Division of Neonatology, Department of Pediatrics, Taipei Medical University, 15Department of Pediatrics and Community Health Sciences, University of Calgary, 16Department of Pediatrics, The Third Affiliated Hospital of Xinxiang Medical University, 17Department of Ultrasound, GE Healthcare, 18Intensive Care Unit, Zhejiang Hospital, 19Collaborative Innovation Center for Maternal and Infant Health Service Application Technology, Quanzhou Medical College, 20Department of Ultrasound, Beijing Children's Hospital Affiliated with Capital Medical University, 21Department of Neonatology and NICU, Tai'an City Central Hospital of Shandong Province, 22Department of Intensive Care Unit, The Second Affiliated Hospital of Heilongjiang University of Chinese Medicine

Lung ultrasound is a noninvasive and valuable tool for bedside evaluation of neonatal lung diseases. However, a relative lack of reference standards, protocols and guidelines may limit its application. Here, we aim to develop a standardized neonatal lung ultrasound diagnostic protocol to be used in clinical decision-making.

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Biology

Laser-Capture Microdissection RNA-Sequencing for Spatial and Temporal Tissue-Specific Gene Expression Analysis in Plants
Lim Chee Liew 1,2, Yan Wang 1,2, Marta Peirats-Llobet 1, Oliver Berkowitz 1,2,3, James Whelan 1,2,3, Mathew G. Lewsey 1,3
1Department of Animal, Plant and Soil Science, AgriBio Building, La Trobe University, 2Australian Research Council Centre of Excellence in Plant Energy Biology, La Trobe University, 3Australian Research Council Research Hub for Medicinal Agriculture, AgriBio Building, La Trobe University

Presented here is a protocol for laser-capture microdissection (LCM) of plant tissues. LCM is a microscopic technique for isolating areas of tissue in a contamination-free manner. The procedure includes tissue fixation, paraffin embedding, sectioning, LCM and RNA extraction. RNA is used in the downstream tissue-specific, temporally resolved analysis of transcriptomes.

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Bioengineering

Lipidico Injection Protocol for Serial Crystallography Measurements at the Australian Synchrotron
Peter Berntsen 1, Rama Sharma 1, Michael Kusel 2, Brian Abbey 1, Connie Darmanin 1
1Australian Research Council Centre of Excellence in Advanced Molecular Imaging, Department of Chemistry and Physics, La Trobe Institute for Molecular Science, La Trobe University, Melbourne, Australia, 3086, 2Kusel Design, Niddrie, Melbourne, Australia, 3042

The goal of this protocol is to demonstrate how to prepare serial crystallography samples for data collection on a high viscous injector, Lipidico, recently commissioned at the Australian synchrotron.

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Environment

Low-Cost Automated Flight Intercept Trap for the Temporal Sub-Sampling of Flying Insects Attracted to Artificial Light at Night
Kylie A. Robert 1, Alicia M. Dimovski 1, Joel A. Robert 2, Stephen R. Griffiths 1
1Department of Ecology, Environment and Evolution, La Trobe University, 2Independent researcher

To study the impacts of artificial light at night (ALAN) on nocturnal flying insects, sampling needs to be confined to nighttime. The protocol describes a low-cost automated flight intercept trap that allows researchers to sample at user-defined periods with increased replication.

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Biology

A Step-By-Step Method to Detect Neutralizing Antibodies Against AAV using a Colorimetric Cell-Based Assay
Sebastian Bass-Stringer 1,2, Colleen J. Thomas 2,3, Clive N. May 3, Paul Gregorevic 4, Kate L. Weeks 1,5,6, Julie R. McMullen 1,2,5,6,7
1Baker Heart and Diabetes Institute, 2Department of Physiology, Anatomy and Microbiology, La Trobe University, 3Florey Institute of Neuroscience and Mental Health, University of Melbourne, 4Department of Physiology, Centre for Muscle Research (CMR), The University of Melbourne, 5Department of Diabetes, Central Clinical School, Monash University, 6Baker Department of Cardiometabolic Health, The University of Melbourne, 7Department of Physiology and Department of Medicine Alfred Hospital, Monash University

A comprehensive laboratory protocol and analysis workflow are described for a rapid, cost-effective, and straightforward colorimetric cell-based assay to detect neutralizing elements against AAV6.

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Medicine

Using a Combination of Indirect Calorimetry, Infrared Thermography, and Blood Glucose Levels to Measure Brown Adipose Tissue Thermogenesis in Humans
Lachlan Van Schaik 1,3, Christine Kettle 1, Rod A. Green 1, Helen R. Irving 1, Joseph A. Rathner 1,2
1La Trobe Institute for Molecular Science, Department of Rural Clinical Sciences, La Trobe University, 2School of Biomedical Sciences, Department of Physiology, The University of Melbourne, 3Melbourne Medical School, Department of Rural Health, The University of Melbourne

Here, we present a protocol to quantify the physiological significance of the impact of brown adipose tissue (BAT) activity on human metabolism. This is achieved by combining carbohydrate loading and indirect calorimetry with measurements of supraclavicular changes in temperature. This novel approach can help develop a pharmacological target for BAT thermogenesis in humans.

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Behavior

Usability Evaluation of Augmented Reality: A Neuro-Information-Systems Study
Jun Wu *1, Di Zhang *1, Tao Liu 2,3,4, Helen Hong Yang 5, Yi Wang 1, Huili Yao 1, Shinan Zhao 1
1Economics and Management School, Jiangsu University of Science and Technology, 2School of Health, Fujian Medical University, 3School of Management, Shanghai University, 4School of Management, Zhejiang University, 5Business School, La Trobe University

This study presents an experimental paradigm for a usability test combining subjective and objective evaluations. The objective evaluation adopted Neuro-Information-Systems (NeuroIS) methods, and the subjective evaluation adopted a usability questionnaire and a NASA-Task Load Index (NASA-TLX) scale.

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Biochemistry

Fiber Type Identification of Human Skeletal Muscle
Heidy K. Latchman *1, Stefan G. Wette *1, Dion J. Ellul 1, Robyn M. Murphy 1, Noni T. Frankenberg 1
1Department of Biochemistry and Chemistry, La Trobe Institute for Molecular Science, School of Agriculture, Biomedicine and Environment, La Trobe University

This protocol demonstrates single-fiber isolation from freeze-dried human skeletal muscle and fiber-type classification according to Myosin heavy chain (MHC) isoform using the dot blotting technique. Identified MHC I and II fiber samples can then be further analyzed for fiber type-specific differences in protein expression using western blotting.

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