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University of Victoria

14 ARTICLES PUBLISHED IN JoVE

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Neuroscience

The Vermicelli and Capellini Handling Tests: Simple quantitative measures of dexterous forepaw function in rats and mice
Kelly A. Tennant 1, Aaron L. Asay 2, Rachel P. Allred 3, Angela R. Ozburn 4, Jeffrey A. Kleim 5, Theresa A. Jones 1,2
1Institute for Neuroscience, University of Texas at Austin, 2Department of Psychology, University of Texas at Austin, 3Department of Neurology, University of Florida, 4Department of Psychiatry, University of Texas Southwestern Medical Center, 5Department of Neuroscience, McKnight Brain Institute, University of Florida

The Vermicelli and Capellini Handling Tests of forepaw dexterity take advantage of the natural inclination of rodents to manipulate food items using skillful forepaw and digit movements. Animals are videotaped while handling short strands of uncooked dry pasta. Slow motion video playback allows for the quantification of forepaw adjustments.

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Biology

Imaging Glycans in Zebrafish Embryos by Metabolic Labeling and Bioorthogonal Click Chemistry
Hao Jiang 1, Lei Feng 1, David Soriano del Amo 1, Ronald D. Seidel III 2, Florence Marlow 3, Peng Wu 1
1Department of Biochemistry, Albert Einstein College of Medicine, Yeshiva University, 2Macromolecular Therapeutics Development Facility, Albert Einstein College of Medicine, Yeshiva University, 3Developmental and Molecular Biology, Albert Einstein College of Medicine, Yeshiva University

A click-chemistry based method that allows for the rapid, noninvasive, and robust labeling of alkyne-tagged glycans in zebrafish embryos is described. Fucosylated glycans in the enveloping layer of zebrafish embryos in the late gastrulation stage were imaged in this study.

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Biology

Quantification of Proteins Using Peptide Immunoaffinity Enrichment Coupled with Mass Spectrometry
Lei Zhao *1, Jeffrey R. Whiteaker *1, Matthew E. Pope 2, Eric Kuhn 3, Angela Jackson 4, N. Leigh Anderson 5, Terry W. Pearson 2, Steven A. Carr 3, Amanda G. Paulovich 1
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.

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Engineering

Optical Trapping of Nanoparticles
Jarrah Bergeron 1, Ana Zehtabi-Oskuie 1, Saeedeh Ghaffari 1, Yuanjie Pang 1, Reuven Gordon 1
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.

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Biology

Dithranol as a Matrix for Matrix Assisted Laser Desorption/Ionization Imaging on a Fourier Transform Ion Cyclotron Resonance Mass Spectrometer
Cuong H. Le 1,2, Jun Han 1, Christoph H. Borchers 1,2
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.

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Engineering

Implementation of a Reference Interferometer for Nanodetection
Serge Vincent *1, Wenyan Yu *1, Tao Lu 1
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.

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Behavior

Compensatory Limb Use and Behavioral Assessment of Motor Skill Learning Following Sensorimotor Cortex Injury in a Mouse Model of Ischemic Stroke
Abigail L. Kerr 1, Kelly A. Tennant 2
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.

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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. Morrison 1, Qi Yuan 1
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.

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Biology

Rapid Identification of Chemical Genetic Interactions in Saccharomyces cerevisiae
David Dilworth 1, Christopher J. Nelson 1
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.

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Chemistry

Peptide and Protein Quantification Using Automated Immuno-MALDI (iMALDI)
Huiyan Li 1, Robert Popp 1, Bjorn Frohlich 1, Michael X. Chen 2, Christoph H. Borchers 1,3,4,5
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.

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Engineering

Plasma-Assisted Molecular Beam Epitaxy Growth of Mg3N2 and Zn3N2 Thin Films
Peng Wu 1, Thomas Tiedje 1
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.

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Biochemistry

Isolation of Histone from Sorghum Leaf Tissue for Top Down Mass Spectrometry Profiling of Potential Epigenetic Markers
Mowei Zhou 1, Shadan H. Abdali 1, David Dilworth 2, Lifeng Liu 2, Benjamin Cole 2, Neha Malhan 1, Amir H. Ahkami 1, Tanya E. Winkler 1, Joy Hollingsworth 3, Julie Sievert 3, Jeff Dahlberg 3, Robert Hutmacher 4,5, Mary Madera 6, Judith A. Owiti 6, Kim K. Hixson 1, Peggy G. Lemaux 6, Christer Jansson 1, Ljiljana Paša-Tolić 1
1Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, 2DOE-Joint Genome Institute, Lawrence Berkeley Laboratory, 3Kearney Agricultural Research and Extension Center, University of California Agriculture and Natural Resources, 4West Side Research and Extension Center, University of California, 5Department of Plant Sciences, University of California, Davis, 6Department of Plant and Microbial Biology, University of California, Berkeley

The protocol has been developed to effectively extract intact histones from sorghum leaf materials for profiling of histone post-translational modifications that can serve as potential epigenetic markers to aid engineering drought resistant crops.

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Immunology and Infection

Measuring the pH, Redox Chemistries, and Degradative Capacity of Macropinosomes using Dual-Fluorophore Ratiometric Microscopy
Liam Wilkinson 1,2, Johnathan Canton 2,3
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.

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Neuroscience

Assessing Changes in Synaptic Plasticity Using an Awake Closed-Head Injury Model of Mild Traumatic Brain Injury
Brian R. Christie 1,2, Allyson Gross 1, Annika Willoughby 1, Erin Grafe 1, Justin Brand 1, Emily Bosdachin 1, Hannah M. O. Reid 1, Crystal Acosta 1, Eric Eyolfson 1
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.

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