University of Ottawa

7 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.

Neuroscience

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

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).

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.

Biology

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

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.

JoVE Core

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

Katie L. Pitts¹, Marianne Fenech^1,2

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.

Neuroscience

Dissection of the Transversus Abdominis Muscle for Whole-mount Neuromuscular Junction Analysis

Lyndsay Murray¹, Thomas H Gillingwater², Rashmi Kothary¹

¹Regenerative Medicine Program, Ottawa Hospital Research Institute, ²Centre for Integrative Physiology, University of Edinburgh

In this video we demonstrate a protocol for dissection of the transversus abdominis muscle of the mouse and use immunofluorescence and microscopy to visualize neuromuscular junctions.

Bioengineering

Controlled Microfluidic Environment for Dynamic Investigation of Red Blood Cell Aggregation

Rym Mehri¹, Catherine Mavriplis¹, Marianne Fenech¹

¹Department of Mechanical Engineering, University of Ottawa

The protocol described details an experimental procedure to quantify Red Blood Cell (RBC) aggregates under a controlled and constant shear rate, based on image processing techniques. The goal of this protocol is to relate RBC aggregate sizes to the corresponding shear rate in a controlled microfluidic environment.