University of Victoria View Institution's Website 13 articles published in JoVE Neuroscience Assessing Changes in Synaptic Plasticity Using an Awake Closed-Head Injury Model of Mild Traumatic Brain Injury Brian R. Christie1,2, Allyson Gross1, Annika Willoughby1, Erin Grafe1, Justin Brand1, Emily Bosdachin1, Hannah M. O. Reid1, Crystal Acosta1, Eric Eyolfson1 1Division of Medical Sciences, University of Victoria, 2Island Medical Program, University of British Columbia Here, it is demonstrated how an awake closed-head injury model can be used for examining the effects of repeated mild traumatic brain injury (r-mTBI) on synaptic plasticity in the hippocampus. The model replicates important features of r-mTBI in patients and is used in conjunction with in vitro electrophysiology. Immunology and Infection Measuring the pH, Redox Chemistries, and Degradative Capacity of Macropinosomes using Dual-Fluorophore Ratiometric Microscopy Liam Wilkinson1,2, Johnathan Canton2,3 1Department of Biochemistry & Microbiology, Faculty of Science, University of Victoria, 2Department of Comparative Biology and Experimental Medicine, Faculty of Veterinary Science, University of Calgary, 3Calvin, Joan and Phoebe Snyder Institute for Chronic Diseases, University of Calgary We describe protocols for measuring pH, oxidative events, and protein digestion in individual macropinosomes in live cells. An emphasis is placed on dual-fluorophore ratiometric microscopy and the advantages it offers over population-based techniques. Engineering Plasma-Assisted Molecular Beam Epitaxy Growth of Mg3N2 and Zn3N2 Thin Films Peng Wu1, Thomas Tiedje1 1Department of Electrical and Computer Engineering, University of Victoria This article describes the growth of epitaxial films of Mg3N2 and Zn3N2 on MgO substrates by plasma-assisted molecular beam epitaxy with N2 gas as the nitrogen source and optical growth monitoring. Chemistry Peptide and Protein Quantification Using Automated Immuno-MALDI (iMALDI) Huiyan Li1, Robert Popp1, Bjorn Frohlich1, Michael X. Chen2, Christoph H. Borchers1,3,4,5 1University of Victoria-Genome BC Proteomics Centre, 2Jewish General Hospital, McGill University, 3Dept of Biochemistry and Microbiology, University of Victoria, 4Proteomics Centre, Segal Cancer Centre, Lady Davis Institute, Jewish General Hospital, McGill University, 5Gerald Bronfman Department of Oncology, Jewish General Hospital A protocol for the protein quantification in complex biological fluids using automated immuno-MALDI (iMALDI) technology is presented. Biology Rapid Identification of Chemical Genetic Interactions in Saccharomyces cerevisiae David Dilworth1, Christopher J. Nelson1 1Department of Biochemistry and Microbiology, University of Victoria Here we present a cost-effective method for defining chemical-genetic interactions in budding yeast. The approach is built on fundamental techniques in yeast molecular biology and is well suited for the mechanistic interrogation of small to medium collections of chemicals and other media environments. Neuroscience A Lateralized Odor Learning Model in Neonatal Rats for Dissecting Neural Circuitry Underpinning Memory Formation Christine J. Fontaine*1,2, Bandhan Mukherjee*1, Gillian L. Morrison1, Qi Yuan1 1Division of Biomedical Sciences, Faculty of Medicine, Memorial University, 2Division of Medical Sciences, University of Victoria This protocol introduces lateralized early odor preference learning in rats using acute single naris occlusion. Lateralized learning permits the examination of behavioral outcomes and underpinning biological mechanisms within the same animals, reducing variance induced by between-animal designs. This protocol can be used to investigate molecular mechanisms underpinning early odor learning. Behavior Compensatory Limb Use and Behavioral Assessment of Motor Skill Learning Following Sensorimotor Cortex Injury in a Mouse Model of Ischemic Stroke Abigail L. Kerr1, Kelly A. Tennant2 1Psychology Department, Illinois Wesleyan University, 2Division of Medical Sciences, University of Victoria Mouse models have become increasingly popular in studies of behavioral neuroscience. As models advance, it is important to develop sensitive behavioral measures specific to the mouse. This protocol describes the Pasta Matrix Reaching Task, which is a skilled motor task for use in mouse models of stroke. Engineering Implementation of a Reference Interferometer for Nanodetection Serge Vincent*1, Wenyan Yu*1, Tao Lu1 1Department of Electrical and Computer Engineering, University of Victoria A reference interferometer technique, which is designed to remove undesirable laser jitter noise for nanodetection, is utilized for probing an ultra-high quality factor microcavity. Instructions for assembly, setup, and data acquisition are provided, alongside the measurement process for specifying the cavity quality factor. Biology Dithranol as a Matrix for Matrix Assisted Laser Desorption/Ionization Imaging on a Fourier Transform Ion Cyclotron Resonance Mass Spectrometer Cuong H. Le1,2, Jun Han1, Christoph H. Borchers1,2 1University of Victoria-Genome BC Proteomics Centre, University of Victoria, 2Department of Biochemistry and Microbiology, University of Victoria Dithranol (DT; 1,8-dihydroxy-9,10-dihydroanthracen-9-one) has previously been reported as a MALDI matrix for tissue imaging of small molecules; protocols for the use of DT for the MALDI imaging of endogenous lipids on the surface of tissue sections by positive-ion MALDI-MS on an ultrahigh-resolution quadrupole-FTICR instrument are provided here. Engineering Optical Trapping of Nanoparticles Jarrah Bergeron1, Ana Zehtabi-Oskuie1, Saeedeh Ghaffari1, Yuanjie Pang1, Reuven Gordon1 1Electrical and Computer Engineering, University of Victoria The following setup approach details low power optical trapping of dielectric nanoparticles using a double-nanohole in metal film. Bioengineering Preparation of 3D Fibrin Scaffolds for Stem Cell Culture Applications Kathleen Kolehmainen1, Stephanie M. Willerth2 1Department of Biology, University of Victoria, 2Department of Mechanical Engineering, Division of Medical Sciences, University of Victoria This work details the preparation of 3D fibrin scaffolds for culturing and differentiating plutipotent stem cells. Such scaffolds can be used to screen the effects of various biological compounds on stem cell behavior as well as modified to contain drug delivery systems. Neuroscience Spectral Confocal Imaging of Fluorescently tagged Nicotinic Receptors in Knock-in Mice with Chronic Nicotine Administration Anthony Renda1, Raad Nashmi1 1Department of Biology, University of Victoria We have developed a novel technique of quantifying nicotinic acetylcholine receptor changes within subcellular regions of specific subtypes of CNS neurons to better understand the mechanisms of nicotine addiction by using a combination of approaches including fluorescent protein tagging of the receptor using the knock-in approach and spectral confocal imaging. Biology Quantification of Proteins Using Peptide Immunoaffinity Enrichment Coupled with Mass Spectrometry Lei Zhao*1, Jeffrey R. Whiteaker*1, Matthew E. Pope2, Eric Kuhn3, Angela Jackson4, N. Leigh Anderson5, Terry W. Pearson2, Steven A. Carr3, Amanda G. Paulovich1 1Clinical Research Division, Fred Hutchinson Cancer Research Center - FHCRC, 2Department of Biochemistry and Microbiology, University of Victoria, 3Broad Institute of MIT and Harvard, 4Genome BC Proteomics Centre, University of Victoria, 5Plasma Proteome Institute Stable Isotope Standards and Capture by Anti-Peptide Antibodies (SISCAPA) couples affinity enrichment of peptides with stable isotope dilution mass spectrometry (MRM-MS) to provide quantitative measurement of peptides as surrogates for their respective proteins. Here we describe the protocol using magnetic particles in a partially automated format.