JoVE Logo

Sign In

University of Calgary

9 ARTICLES PUBLISHED IN JoVE

image

Neuroscience

Efficient Gene Delivery into Multiple CNS Territories Using In Utero Electroporation
Rajiv Dixit 1, Fuqu Lu 2, Robert Cantrup 1, Nicole Gruenig 1,2, Lisa Marie Langevin 1, Deborah M. Kurrasch 2, Carol Schuurmans 1
1Department of Biochemistry and Molecular Biology, Hotchkiss Brain Institute, Alberta Children’s Hospital Research Institute, University of Calgary , 2Department of Medical Genetics, Alberta Children’s Hospital Research Institute, Hotchkiss Brain Institute, University of Calgary

In utero electroporation allows for rapid gene delivery in a spatially- and temporally-controlled manner in the developing central nervous system (CNS). Here we describe a highly adaptable in utero electroporation protocol that can be used to deliver expression constructs into multiple embryonic CNS domains, including the telencephalon, diencephalon and retina.

image

Biology

Procedures for Rat in situ Skeletal Muscle Contractile Properties
Brian R. MacIntosh 1, Shane P. Esau 1, R. John Holash 1, Jared R. Fletcher 1
1Faculty of Kinesiology, University of Calgary

This video demonstrates the surgical preparation and procedures needed to study the contractile responses of the rat medial gastrocnemius muscle preparation in situ. This preparation allows measurement of skeletal muscle contractile properties under physiological conditions. The animal is anesthetized and the muscle is separated from surrounding tissue at its distal end. The Achilles tendon is attached to a force transducer, allowing measurement of the muscle’s contractile response at 37 degrees C with an intact circulation.

image

Neuroscience

Culturing and Electrophysiology of Cells on NRCC Patch-clamp Chips
Christophe Py 1, Marzia Martina 2, Robert Monette 2, Tanya Comas 2, Mike W. Denhoff 1, Collin Luk 3, Naweed I. Syed 3, Geoff Mealing 2
1Institute for Microstructural Sciences, National Research Council of Canada, 2Institute for Biological Sciences, National Research Council of Canada, 3Hotchkiss Brain Institute, University of Calgary

We show how planar patch-clamp chips fabricated at the National Research Council of Canada are sterilized, primed, loaded with medium, plated with cells, and used for electrophysiological recordings.

image

Biology

Germ Cell Transplantation and Testis Tissue Xenografting in Mice
Lin Tang *1, Jose Rafael Rodriguez-Sosa *1, Ina Dobrinski 1
1Department of Comparative Biology and Experimental Medicine, University of Calgary

Protocols for germ cell transplantation and testis tissue xenografting are described. Germ cell transplantation results in donor-derived spermatogenesis in recipient testes and represents a functional reconstitution assay for identification of spermatogonial stem cells (SSCs). Testis tissue xenografting reproduces testis development and spermatogenesis of various donor species in recipient mice.

image

Immunology and Infection

Isolation of Human Umbilical Vein Endothelial Cells and Their Use in the Study of Neutrophil Transmigration Under Flow Conditions
Anutosh Ganguly 1, Hong Zhang 1, Ritu Sharma 1, Sean Parsons 1, Kamala D. Patel 1
1Department of Physiology and Pharmacology, University of Calgary

This article first describes a procedure for isolating human endothelial cells from umbilical veins and then shows how to use these cells to examine neutrophil transmigration under flow conditions. By using a low-volume flow chamber made from a polymer with the optical characteristics of glass, live-cell fluorescent imaging of rare cell populations is also possible.

image

Neuroscience

Wholemount Immunohistochemistry for Revealing Complex Brain Topography
Joshua J. White 1, Stacey L. Reeber 1, Richard Hawkes 2, Roy V. Sillitoe 1
1Department of Neuroscience, Albert Einstein College of Medicine, Yeshiva University , 2Department of Cell Biology and Anatomy and the Hotchkiss Brain Institute, Faculty of Medicine, University of Calgary

Neural circuits are topographically organized into functional compartments with specific molecular profiles. Here, we provide the practical and technical steps for revealing global brain topography using a versatile wholemount immunohistochemical staining approach. We demonstrate the utility of the method using the well-understood cytoarchitecture and circuitry of cerebellum.

image

Medicine

Detection of Architectural Distortion in Prior Mammograms via Analysis of Oriented Patterns
Rangaraj M. Rangayyan 1,2, Shantanu Banik 1, J.E. Leo Desautels 1
1Department of Electrical and Computer Engineering, Schulich School of Engineering, University of Calgary , 2Department of Radiology, University of Calgary

We demonstrate methods for the detection of architectural distortion in prior mammograms. Oriented structures are analyzed using Gabor filters and phase portraits to detect sites of radiating tissue patterns. Each site is characterized and classified using measures to represent spiculating patterns. The methods should assist in the detection of breast cancer.

image

Medicine

Generation of Porcine Testicular Organoids with Testis Specific Architecture using Microwell Culture
Sadman Sakib 1, Yang Yu 3,4, Anna Voigt 2, Mark Ungrin 2,3,4, Ina Dobrinski 1,2
1Biochemistry and Molecular Biology, University of Calgary, 2Comparative Biology and Experimental Medicine, University of Calgary, 3Biomedical Engineering Graduate Program, University of Calgary, 4Alberta Diabetes Institute, University of Alberta

Here, we present a protocol for the reproducible generation of porcine testicular organoids with testis specific tissue architecture using the commercially available microwell culture system.

image

Immunology and Infection

Intravital Widefield Fluorescence Microscopy of Pulmonary Microcirculation in Experimental Acute Lung Injury Using a Vacuum-Stabilized Imaging System
Stefan Hall 1, Sufyan Faridi 2, Irene Euodia 2,3, Sophie Tanner 4, Andrew Krzysztof Chojnacki 5, Kamala D. Patel 6,7, Juan Zhou 2,4, Christian Lehmann 1,2,4,8
1Department of Physiology and Biophysics, Dalhousie University, 2Department of Microbiology and Immunology, Dalhousie University, 3Department of Neuroscience, Dalhousie University, 4Department of Anesthesia, Pain Management, and Perioperative Medicine, Dalhousie University, 5Live Cell Imaging Center, University of Calgary, 6Department of Physiology and Pharmacology, University of Calgary, 7Department of Biochemistry and Molecular Biology, University of Calgary, 8Department of Pharmacology, Dalhousie University

Intravital fluorescence microscopy can be utilized to study leukocyte-endothelial interactions and capillary perfusion in real-time. This protocol describes methods to image and quantify these parameters in the pulmonary microcirculation using a vacuum-stabilized lung imaging system.

JoVE Logo

Privacy

Terms of Use

Policies

Research

Education

ABOUT JoVE

Copyright © 2024 MyJoVE Corporation. All rights reserved