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Carleton University

5 ARTICLES PUBLISHED IN JoVE

Neuroscience

Localizing Protein in 3D Neural Stem Cell Culture: a Hybrid Visualization Methodology

Sophie Imbeault^*1, Nicolas Valenzuela^*2, Stephen Fai², Steffany A.L. Bennett¹

¹Neural Regeneration Laboratory and Ottawa Institute of Systems Biology, Department of Biochemistry, Microbiology and Immunology, University of Ottawa, ²Carleton Immersive Media Studio, Azrieli School of Architecture and Urbanism, Carleton University

Here, we describe how to produce, expand, and immunolabel postnatal hippocampal neural progenitor cells (NPCs) in three-dimensional (3D) culture. Next, using hybrid visualization technologies, we demonstrate how digital images of immunolabelled cryosections can be used to reconstruct and map the spatial position of immunopositive cells throughout the entire 3D neurosphere.

Biology

Performing Vaginal Lavage, Crystal Violet Staining, and Vaginal Cytological Evaluation for Mouse Estrous Cycle Staging Identification

Ashleigh C. McLean^1,2,3, Nicolas Valenzuela^3,4, Stephen Fai^3,4, Steffany A.L. Bennett^1,3

¹Department of Biochemistry, Microbiology and Immunology, Neural Regeneration Laboratory and Ottawa Institute of Systems Biology, ²Department of Cellular and Molecular Medicine, University of Ottawa , ³CIHR Program in Neurodegenerative Lipidomics, University of Ottawa , ⁴Carleton Immersive Media Studio, Azrieli School of Architecture and Urbanism

Here, we describe how to identify the stage of the murine reproductive (proestrus, estrus, metestrus, or diestrus) by simple, non-invasive collection and cytological assessment of vaginal smear samples. We further describe how vaginal cytology reflects circulating hormonal levels underlying transition through the murine reproductive cycle.

Environment

Split Point Analysis and Uncertainty Quantification of Thermal-Optical Organic/Elemental Carbon Measurements

Bradley M. Conrad¹, Matthew R. Johnson¹

¹Energy and Emissions Research Laboratory, NSERC FlareNet Strategic Network, Department of Mechanical and Aerospace Engineering, Carleton University

This article presents a protocol and software tool for the quantification of uncertainties in the calibration and data analysis of a semi-continuous thermal-optical organic/elemental carbon analyzer.

Immunology and Infection

Generation of Null Mutants to Elucidate the Role of Bacterial Glycosyltransferases in Bacterial Motility

Juan M. Tomás¹, Kelly M. Fulton^2,3, Susan M. Twine^2,4, Susana Merino¹

¹Departamento de Genética, Microbiología y Estadística, Universidad de Barcelona, ²Human Health Therapeutics Research Centre, National Research Council Canada, ³Department of Chemistry, Carleton University, ⁴Department of Biology, Carleton University

Here, we describe the construction of null mutants of Aeromonas in specific glycosyltransferases or regions containing glycosyltransferases, the motility assays, and flagella purification performed to establish the involvement and function of their encoded enzymes in the biosynthesis of a glycan, as well as the role of this glycan in bacterial pathogenesis.

JoVE Journal

Modifying Levels of Maternal Dietary Folic Acid or Choline to Study the Impact of Deficiencies on Offspring Health Outcomes

Mitra Esfandiarei^1,2,3, Teodoro Bottiglieri⁴, Nafisa M. Jadavji^5,6,7

¹Department of Biomedical Sciences, Midwestern University, ²Department of Basic Medical Sciences, College of Medicine Phoenix, University of Arizona, ³Department of Anesthesiology, Pharmacology, & Therapeutics, Faculty of Medicine, University of British Columbia, ⁴Center of Metabolomics, Institute of Metabolic Disease, Baylor Scott & White Research Institute, ⁵Department of Physiology, Southern Illinois University, ⁶Department of Child Health, College of Medicine Phoenix, University of Arizona, ⁷Department of Neuroscience, Carleton University

This protocol describes the method for reducing dietary intake of folic acid or choline in female mice prior to pregnancy with the objective of investigating the impact of maternal diet on offspring health outcomes.