University of Ottawa

7 articles published in JoVE

Micro-particle Image Velocimetry for Velocity Profile Measurements of Micro Blood Flows

JoVE 50314 4/25/2013

¹Department of Chemical and Biological Engineering, University of Ottawa, ²Department of Mechanical Engineering, University of Ottawa

Micro-particle image velocimetry (μPIV) is used to visualize paired images of micro particles seeded in blood flows which are cross-correlated to give an accurate velocity profile. Shear rate, maximum velocity, velocity profile shape, and flow rate, each of which has clinical applications, can be derived from these measurements.

Isolation and Culture of Individual Myofibers and their Satellite Cells from Adult Skeletal Muscle

JoVE 50074 3/22/2013

Alessandra Pasut^1,2, Andrew E. Jones^1,2, Michael A. Rudnicki^1,2

¹Sprott Center for Stem Cell Research, Ottawa Hospital Research Institute, ²Department of Cellular and Molecular Medicine, University of Ottawa

Isolation and culture of myofibers is the gold standard in vitro system to study the transition of satellite cells through quiescence, activation and differentiation. Importantly, the single myofiber culture system preserves the myofiber/stem cell association, which is an essential component of the muscle stem cell niche.

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

JoVE 4389 9/15/2012

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.

Derivation of Enriched Oligodendrocyte Cultures and Oligodendrocyte/Neuron Myelinating Co-cultures from Post-natal Murine Tissues

JoVE 3324 8/21/2011

Ryan W. O'Meara^1,2, Scott D. Ryan¹, Holly Colognato³, Rashmi Kothary^1,2,4

¹Regenerative Medicine Program, Ottawa Hospital Research Institute, ²Department of Cellular and Molecular Medicine, University of Ottawa, ³Department of Pharmacological Sciences, Stony Brook University, ⁴Department of Medicine, University of Ottawa

This article describes methods to derive enriched populations of murine oligodendrocyte precursor cells (OPCs) in primary culture, which differentiate to produce mature oligodendrocytes (OLs). In addition, this report describes techniques to produce murine myelinating co-cultures by seeding mouse OPCs onto a neurite bed of mouse dorsal root ganglion neurons (DRGNs).

Lentiviral-mediated Knockdown During Ex Vivo Erythropoiesis of Human Hematopoietic Stem Cells

JoVE 2813 7/16/2011

Carmen G. Palii¹, Roya Pasha¹, Marjorie Brand^1,2

¹The Sprott Center for Stem Cell Research, Regenerative Medicine Program, Ottawa Hospital Research Institute, ²Department of Cellular and Molecular Medicine, University of Ottawa

An ex vivo protocol to generate mature human red blood cells from hematopoietic stem/progenitors is described. Additionally we describe an efficient lentiviral-delivery method to knockdown the transcription factor TAL1 in primary erythroid cells. The efficiency of lentivirus mediated gene delivery is demonstrated using GFP expressing viruses.

Quantitative Analyses of all Influenza Type A Viral Hemagglutinins and Neuraminidases using Universal Antibodies in Simple Slot Blot Assays

JoVE 2784 4/04/2011

Caroline Gravel*¹, Changgui Li*², Junzhi Wang², Anwar M Hashem^1,3,4, Bozena Jaentschke¹, Gary Van Domselaar⁵, Runtao He⁵, Xuguang Li^1,3

¹Centre for Vaccine Evaluation, Biologics and Genetic Therapies Directorate, HPFB, Health canada, ²National Institute for the Control of Pharmaceutical and Biological Products, The State Food and Drug Administration, Beijing, ³Department of Biochemistry, Microbiology and Immunology, University of Ottawa, ⁴Microbiology Department, Faculty of Medicine, King Abdulaziz University, ⁵National Microbiology Laboratory, Public Health Agency of Canada

A simple slot blot method was developed for the quantification of influenza viral hemagglutinin and neuraminidase using universal antibodies targeting their most conserved sequences identified through bioinformatics analyses. This innovative approach may provide a useful alternative to quantitative determination of all viral hemagglutinin and neuraminidase.

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

JoVE 2483 12/19/2010

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.