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Concordia University

20 ARTICLES PUBLISHED IN JoVE

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Biology

Dissection of Saccharomyces Cerevisiae Asci
Audrey Morin 1, Adrian W. Moores 1, Michael Sacher 1
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.

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Biology

Seawater Sampling and Collection
Elena Zaikova 1, Alyse Hawley 1, David A. Walsh 1, Steven J. Hallam 1
1Department of Microbiology and Immunology, University of British Columbia - UBC

This video documents methods for collecting coastal marine water samples and processing them for various downstream applications including biomass concentration, nucleic acid purification, cell abundance, nutrient and trace gas analyses.

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Biology

Large Volume (20L+) Filtration of Coastal Seawater Samples
David A. Walsh 1, Elena Zaikova 1, Steven J. Hallam 1
1Department of Microbiology and Immunology, University of British Columbia - UBC

This video documents large volume (≥20 L) filtration of microbial biomass, ranging between 0.22μm and 2.7μm in diameter, from the water column.

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JoVE Core

Small Volume (1-3L) Filtration of Coastal Seawater Samples
David A. Walsh 1, Elena Zaikova 1, Steven J. Hallam 1
1Department of Microbiology and Immunology, University of British Columbia - UBC

This video documents small volume (~1 L) filtration of microbial biomass from the water column.

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Biology

DNA Extraction from 0.22 μM Sterivex Filters and Cesium Chloride Density Gradient Centrifugation
Jody J. Wright 1, Sangwon Lee 1, Elena Zaikova 1, David A. Walsh 1, Steven J. Hallam 1
1Department of Microbiology and Immunology, University of British Columbia - UBC

We describe a method for extraction of high molecular weight genomic DNA from planktonic biomass concentrated on 0.22 μm Sterivex filters, followed by cesium chloride density gradient centrifugation for purification.

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Biology

Purification of Mitochondria from Yeast Cells
Christopher Gregg 1, Pavlo Kyryakov 1, Vladimir I. Titorenko 1
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.

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Biology

A Quantitative Assessment of The Yeast Lipidome using Electrospray Ionization Mass Spectrometry
Simon D. Bourque 1, Vladimir I. Titorenko 1
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.

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Neuroscience

Recording and Analysis of Circadian Rhythms in Running-wheel Activity in Rodents
Michael Verwey 1, Barry Robinson 2, Shimon Amir 2
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.

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Medicine

Using Continuous Data Tracking Technology to Study Exercise Adherence in Pulmonary Rehabilitation
Amanda K. Rizk 1,2,3, Rima Wardini 2,3, Emilie Chan-Thim 1,2,3, Barbara Trutschnigg 3, Amélie Forget 3, Véronique Pepin 2,3
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.

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Behavior

A Procedure to Study the Effect of Prolonged Food Restriction on Heroin Seeking in Abstinent Rats
Firas Sedki 1, Tracey D'Cunha 1, Uri Shalev 1
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.

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Neuroscience

Visualizing Neuroblast Cytokinesis During C. elegans Embryogenesis
Denise Wernike 1, Chloe van Oostende 1, Alisa Piekny 1
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.

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Behavior

A Procedure to Observe Context-induced Renewal of Pavlovian-conditioned Alcohol-seeking Behavior in Rats
Jean-Marie Maddux 1, Franca Lacroix 1, Nadia Chaudhri 1
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.

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Environment

An Aquatic Microbial Metaproteomics Workflow: From Cells to Tryptic Peptides Suitable for Tandem Mass Spectrometry-based Analysis
David Colatriano 1, David A. Walsh 1
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.

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Engineering

A Simple Dewar/Cryostat for Thermally Equilibrating Samples at Known Temperatures for Accurate Cryogenic Luminescence Measurements
Phoebe G. Weaver 1, Devin M. Jagow 1, Cameron M. Portune 1, John W. Kenney, III 1
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.

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Behavior

Eye Tracking During Visually Situated Language Comprehension: Flexibility and Limitations in Uncovering Visual Context Effects
Camilo Rodriguez Ronderos 1, Katja Münster 1, Ernesto Guerra 2, Helene Kreysa 3, Alba Rodríguez 1,4, Julia Kröger 4, Thomas Kluth 4, Michele Burigo 4, Dato Abashidze 5, Eva Nunnemann 4, Pia Knoeferle 1,6,7
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.

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JoVE Core

Operation of the Collaborative Composite Manufacturing (CCM) System
Pengcheng Li 1, Xiaoming Zhang 1, Wenfang Xie 1, Suong Van Hoa 1
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.

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Biochemistry

Quantitative Analysis of the Cellular Lipidome of Saccharomyces Cerevisiae Using Liquid Chromatography Coupled with Tandem Mass Spectrometry
Karamat Mohammad 1, Heng Jiang 2, Md. Israil Hossain 1, Vladimir I. Titorenko 1
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.

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Biochemistry

Quantitative Metabolomics of Saccharomyces Cerevisiae Using Liquid Chromatography Coupled with Tandem Mass Spectrometry
Karamat Mohammad 1, Heng Jiang 2, Vladimir I. Titorenko 1
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.

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Behavior

A Procedure to Study Stress-Induced Relapse of Heroin Seeking after Punishment-Imposed Abstinence
Catarina Borges 1, Jordan Charles 1, Uri Shalev 1
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.

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Biology

Identification of the Genes Involved in Stomatal Development via Epidermal Phenotype Scoring
Pooja Kaushik *1, Kritika Bharti *1, Jin Suk Lee 1
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.

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