Concordia University 17 articles published in JoVE Biology Identification of the Genes Involved in Stomatal Development via Epidermal Phenotype Scoring Pooja Kaushik*1, Kritika Bharti*1, Jin Suk Lee1 1Department of Biology, Concordia University This paper describes two phenotyping methods without the use of epidermal peels to characterize the genes controlling stomatal development. The first method demonstrates how to analyze a stomatal phenotype using a toluidine blue O-stained plant epidermis. The second method describes how to identify stomatal ligands and monitor their biological activities. Behavior A Procedure to Study Stress-Induced Relapse of Heroin Seeking after Punishment-Imposed Abstinence Catarina Borges1, Jordan Charles1, Uri Shalev1 1Department of Psychology, Center for Studies in Behavioral Neurobiology, Concordia University A procedure that demonstrates a robust acute food deprivation-induced relapse to heroin seeking after punishment-imposed abstinence is described. A punishment-imposed abstinence model was successfully implemented using the seek and take chain schedule for heroin self-administration. Heroin-seeking tests are then performed following 24 h of food-deprivation stress. Biochemistry Quantitative Metabolomics of Saccharomyces Cerevisiae Using Liquid Chromatography Coupled with Tandem Mass Spectrometry Karamat Mohammad1, Heng Jiang2, Vladimir I. Titorenko1 1Department of Biology, Concordia University, 2Centre for Biological Applications of Mass Spectrometry, Concordia University We present a protocol for the identification and quantitation of major classes of water-soluble metabolites in the yeast Saccharomyces cerevisiae. The described method is versatile, robust, and sensitive. It allows the separation of structural isomers and stereoisomeric forms of water-soluble metabolites from each other. Biochemistry Quantitative Analysis of the Cellular Lipidome of Saccharomyces Cerevisiae Using Liquid Chromatography Coupled with Tandem Mass Spectrometry Karamat Mohammad1, Heng Jiang2, Md. Israil Hossain1, Vladimir I. Titorenko1 1Department of Biology, Concordia University, 2Department of Chemistry and Biochemistry, Centre for Biological Applications of Mass Spectrometry, Concordia University We present a protocol using liquid chromatography coupled with tandem mass spectrometry to identify and quantify major cellular lipids in Saccharomyces cerevisiae. The described method for a quantitative assessment of major lipid classes within a yeast cell is versatile, robust, and sensitive. Engineering Operation of the Collaborative Composite Manufacturing (CCM) System Pengcheng Li1, Xiaoming Zhang1, Wenfang Xie1, Suong Van Hoa1 1Department of Mechanical, Industrial & Aerospace Engineering, Concordia University A collaborative composite manufacturing system is developed for robotic lay-up of composite laminates using the prepreg tape. The proposed system allows the production of composite laminates with high levels of geometrical complexity. The issues in the path planning, coordination of the robots and control are addressed in the proposed method. Behavior Eye Tracking During Visually Situated Language Comprehension: Flexibility and Limitations in Uncovering Visual Context Effects Camilo Rodriguez Ronderos1, Katja Münster1, Ernesto Guerra2, Helene Kreysa3, Alba Rodríguez1,4, Julia Kröger4, Thomas Kluth4, Michele Burigo4, Dato Abashidze5, Eva Nunnemann4, Pia Knoeferle1,6,7 1Institute of German Studies and Linguistics, Humboldt-Universität zu Berlin, 2Center for Advanced Research in Education – CIAE, Universidad de Chile, 3Institute of Psychology, Friedrich Schiller University Jena, 4Cognitive Interaction Technology Excellence Cluster (CITEC), Faculty of Linguistics and Literature, Bielefeld University, 5Department of Education, Concordia University, 6Berlin School of Mind and Brain, 7Einstein Center for Neurosciences Berlin The present article reviews an eye-tracking methodology for studies on language comprehension. To obtain reliable data, key steps of the protocol must be followed. Among these are the correct set-up of the eye tracker (e.g., ensuring good quality of the eye and head images) and accurate calibration. Engineering A Simple Dewar/Cryostat for Thermally Equilibrating Samples at Known Temperatures for Accurate Cryogenic Luminescence Measurements Phoebe G. Weaver1, Devin M. Jagow1, Cameron M. Portune1, John W. Kenney, III1 1Chemical Physics Laboratory, Concordia University A simple liquid nitrogen Dewar/cryostat apparatus comprised of a small fused silica optical Dewar, a thermocouple, and a charge-coupled device (CCD) spectrograph are described. The experiments for which this Dewar/cryostat is designed require fast sample loading, freezing, and alignment, accurate and stable sample temperatures, and small size/portability. Environment An Aquatic Microbial Metaproteomics Workflow: From Cells to Tryptic Peptides Suitable for Tandem Mass Spectrometry-based Analysis David Colatriano1, David A. Walsh1 1Department of Biology, Concordia University This protocol is for the extraction and concentration of protein and DNA from microbial biomass collected from seawater, followed by the generation of tryptic peptides suitable for tandem mass spectrometry-based proteomic analysis. Immunology and Infection Detecting Glycogen in Peripheral Blood Mononuclear Cells with Periodic Acid Schiff Staining Mahdieh Tabatabaei Shafiei1, Catalina M. Carvajal Gonczi1, Mohammed Samiur Rahman2, Ashley East3, Jonathan François3, Peter J. Darlington1,3 1Department of Biology, Centre for Structural and Functional Genomics, PERFORM Centre, Concordia University, 2Department of Chemistry and Biochemistry, Centre for Structural and Functional Genomics, PERFORM Centre, Concordia University, 3Department of Exercise Science, Centre for Structural and Functional Genomics, PERFORM Centre, Concordia University Periodic acid Schiff staining is a technique that visualizes the polysaccharide content of tissues. This article demonstrates periodic acid Schiff staining protocol adapted for use on peripheral blood mononuclear cells purified from human venous blood. Such samples are enriched for lymphocytes and other white blood cells of the immune system. Behavior A Procedure to Observe Context-induced Renewal of Pavlovian-conditioned Alcohol-seeking Behavior in Rats Jean-Marie Maddux1, Franca Lacroix1, Nadia Chaudhri1 1Department of Psychology, Center for Studies in Behavioral Neurobiology, Concordia University A procedure to study the capacity of an alcohol associated environmental context to trigger the renewal of alcohol-seeking behavior in rats is described. Neuroscience Visualizing Neuroblast Cytokinesis During C. elegans Embryogenesis Denise Wernike1, Chloe van Oostende1, Alisa Piekny1 1Department of Biology, Concordia University This protocol describes how to image dividing cells within a tissue in Caenorhabditis elegans embryos. While several protocols describe how to image cell division in the early embryo, this protocol describes how to image cell division within a developing tissue during mid late embryogenesis. Behavior A Procedure to Study the Effect of Prolonged Food Restriction on Heroin Seeking in Abstinent Rats Firas Sedki1, Tracey D'Cunha1, Uri Shalev1 1Department of Psychology, Center for Studies in Behavioural Neurobiology/Groupe de Recherche en Neurobiologie Comportementale, Concordia University A procedure that allows a demonstration of robust augmentation of drug seeking in food-restricted rats is described. Following heroin self-administration training, rats go through an abstinence period, in a drug-free environment, during which they are mildly food restricted. Drug seeking is then tested in the drug-associated environment. Medicine Using Continuous Data Tracking Technology to Study Exercise Adherence in Pulmonary Rehabilitation Amanda K. Rizk1,2,3, Rima Wardini2,3, Emilie Chan-Thim1,2,3, Barbara Trutschnigg3, Amélie Forget3, Véronique Pepin2,3 1Individualized Program, Concordia University, 2Department of Exercise Science, Concordia University, 3Centre de Recherche, Axe Maladies Chroniques, Hôpital du Sacré-Coeur de Montréal Pulmonary rehabilitation is widely recognized in the management of respiratory diseases. A key component to successful pulmonary rehabilitation is adherence to the recommended exercise training. The purpose of the present protocol is to describe how continuous data tracking technology can be used to precisely measure adherence to a prescribed aerobic training intensity. Neuroscience Recording and Analysis of Circadian Rhythms in Running-wheel Activity in Rodents Michael Verwey1, Barry Robinson2, Shimon Amir2 1Douglas Mental Health University Institute, Department of Psychiatry, McGill University, 2Center for Studies in Behavioral Neurobiology, Department of Psychology, Concordia University Circadian rhythms in voluntary wheel-running activity in mammals are tightly coupled to the molecular oscillations of a master clock in the brain. As such, these daily rhythms in behavior can be used to study the influence of genetic, pharmacological, and environmental factors on the functioning of this circadian clock. Biology Purification of Mitochondria from Yeast Cells Christopher Gregg1, Pavlo Kyryakov1, Vladimir I. Titorenko1 1Department of Biology, Concordia University We describe a rapid and effective method for purification of mitochondria from the yeast Saccharomyces cerevisiae. This method enables the high-yield isolation of pure mitochondria that are essentially free of contamination by other organelles and retain their structural and functional integrity after their purification. Biology A Quantitative Assessment of The Yeast Lipidome using Electrospray Ionization Mass Spectrometry Simon D. Bourque1, Vladimir I. Titorenko1 1Department of Biology, Concordia University We describe a new quantitative lipidomics method for identifying numerous lipid species in yeast using survey-scan electrospray ionization mass spectrometry (ESI/MS). This method exceeds currently available methods for lipid identification and quantification in the ability to resolve various molecular forms of lipids, sensitivity, and speed. Biology Dissection of Saccharomyces Cerevisiae Asci Audrey Morin1, Adrian W. Moores1, Michael Sacher1 1Department of Biology, Concordia University Micromanipulation of yeast cells is needed for meiotic genetic analysis or to select diploid zygotes. These micromanipulations are carried out using the microneedle of a dissection microscope. The microneedle is used to relocate cells and is controlled by a micromanipulator which are available with various degrees of automation.