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University of British Columbia

44 ARTICLES PUBLISHED IN JoVE

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Immunology and Infection

Facilitating the Analysis of Immunological Data with Visual Analytic Techniques
David C. Shih 1,2, Kevin C. Ho 1, Kyle M. Melnick 3, Ronald A. Rensink 2,3, Tobias R. Kollmann 1, Edgardo S. Fortuno III 1
1Department of Paediatrics, Division of Infectious and Immunological Diseases, Child and Family Research Institute, University of British Columbia, 2Department of Computer Science, University of British Columbia, 3Department of Psychology, University of British Columbia

Visual analytics (VA) is a new approach of analyzing data interactively. In this video, we discuss the data overload problem brought on by high-throughput biological experiments, and propose VA as a solution to such problem. The video demonstrates analysis within and between immunological datasets using a VA tool called Tableau.

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Biology

Purification of Progenitors from Skeletal Muscle
Lin Yi 1, Fabio Rossi 1
1The Biomedical Research Centre, University of British Columbia

Method for the enzymatic dissociation, surface labeling and purification by flow cytometry of fibro/adipogenic and myogenic progenitors from murine skeletal muscle.

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Immunology and Infection

Trichuris muris Infection: A Model of Type 2 Immunity and Inflammation in the Gut
Frann Antignano *1, Sarah C. Mullaly *1, Kyle Burrows 1, Colby Zaph 1,2
1The Biomedical Research Centre, University of British Columbia, 2Department of Pathology and Laboratory Medicine, University of British Columbia

Trichuris muris infection is an intestinal model of Th2 immunity where resistant mice generate a protective Th2 response and susceptible mice generate a pathological Th1 response.

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Neuroscience

Morris Water Maze Test for Learning and Memory Deficits in Alzheimer's Disease Model Mice
Kelley Bromley-Brits *1, Yu Deng *1, Weihong Song 1
1Department of Psychiatry, Brain Research Center, University of British Columbia

The Morris Water Maze is a behavioral task to test hippocampal-dependent learning and memory. It has been widely used in the study of neurobiology, neuropharmacology and neurocognitive disorders in rodent models.

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Medicine

Minimally Invasive Establishment of Murine Orthotopic Bladder Xenografts
Wolfgang Jäger 1, Igor Moskalev 1, Claudia Janssen 1, Tetsutaro Hayashi 1, Killian M. Gust 1, Shannon Awrey 1, Peter C. Black 1
1Department of Urologic Sciences, University of British Columbia

The established technique to inoculate primary invasive orthotopic bladder cancer xenografts requires laparotomy and mobilization of the bladder. This procedure inflicts significant morbidity on the mice, is technically challenging and time-consuming. We therefore developed a high-precision, percutaneous approach utilizing ultrasound guidance.

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Behavior

Operant Procedures for Assessing Behavioral Flexibility in Rats
Anne Marie Brady 1, Stan B. Floresco 2
1Department of Psychology, St. Mary's College of Maryland, 2Department of Psychology, University of British Columbia

The ability to assess executive functions such as behavioral flexibility in rats is useful for investigating the neurobiology of cognition in both intact animals and disease models. Here we describe automated tasks for assessing strategy shifting and reversal learning, which are particularly sensitive to disruptions in prefrontal cortical networks.

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Medicine

Assessing Transmissible Spongiform Encephalopathy Species Barriers with an In Vitro Prion Protein Conversion Assay
Christopher J. Johnson 1, Christina M. Carlson 2, Aaron R. Morawski 3, Alyson Manthei 4, Neil R. Cashman 5
1USGS National Wildlife Health Center, 2Department of Soil Science, University of Wisconsin–Madison, 3Laboratory of Immunology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 4Merial Veterinary Scholars Program, School of Veterinary Medicine, University of Wisconsin–Madison, 5Department of Neurology, University of British Columbia

Measuring the barrier to the interspecies transmission of prion diseases is challenging and typically involves animal challenges or biochemical assays. Here, we present an in vitro prion protein conversion assay with the ability to predict species barriers.

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Medicine

Busulfan as a Myelosuppressive Agent for Generating Stable High-level Bone Marrow Chimerism in Mice
Kyle Peake 1, John Manning 1, Coral-Ann Lewis 1,2, Christine Barr 1, Fabio Rossi 2, Charles Krieger 1,3
1Department of Biomedical Physiology and Kinesiology, Simon Fraser University, 2The Biomedical Research Centre, University of British Columbia, 3Division of Neurology, Department of Medicine, Neuromuscular Disease Unit, VHHSC

We describe a protocol whereby busulfan conditioning permits the bone marrow of a recipient mouse to be replaced with bone marrow cells from donor mice ubiquitously expressing green fluorescent protein, in the absence of irradiation. This technique is useful to study bone marrow cell accumulation in the central nervous system.

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Education

Visualization of High Speed Liquid Jet Impaction on a Moving Surface
Yuchen Guo 1, Sheldon Green 1
1Department of Mechanical Engineering, University of British Columbia

Two experimental devices for examining liquid jet impingement on a high-speed moving surface are described: an air cannon device and a spinning disk device. The apparatuses are used to determine optimal approaches to the application of liquid friction modifier (LFM) onto rail tracks for top-of-rail friction control.

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Biology

Transfecting RAW264.7 Cells with a Luciferase Reporter Gene
Sylvia T. Cheung 1,2,3, Soroush Shakibakho 1,2, Eva Y. So 1,2, Alice L-F Mui 1,2,3
1Immunity and Infection Research Centre, Vancouver Costal Health Research Institute, 2Department of Surgery, University of British Columbia, 3Department of Biochemistry and Molecular Biology, University of British Columbia

Transfection into the macrophage cell line, RAW264.7, is difficult due to the cell’s natural response against foreign materials. We described here a gentle yet robust procedure for transfecting luciferase reporter genes into RAW264.7 cells.

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Biology

Measurement of Calcium Fluctuations Within the Sarcoplasmic Reticulum of Cultured Smooth Muscle Cells Using FRET-based Confocal Imaging
Gabriela Ziomek 1, Cornelis van Breemen 1, Mitra Esfandiarei 1,2
1Department of Anaesthesiology, Pharmacology, and Therapeutics, University of British Columbia, 2Department of Biomedical Sciences, Midwestern University

Currently, most available calcium indicators are used to quantify cytoplasmic calcium transients as indirect measures of calcium released from the sarcoplasmic reticulum in cultured smooth muscle cells. This protocol describes the use of a specific FRET-based indicator that allows direct measurement of calcium signals within the sarcoplasmic reticulum lumen.

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Immunology and Infection

Generation and Identification of GM-CSF Derived Alveolar-like Macrophages and Dendritic Cells From Mouse Bone Marrow
Yifei Dong 1, Arif A. Arif 1, Grace F. T. Poon 1, Blair Hardman 1, Manisha Dosanjh 1, Pauline Johnson 1
1Microbiology and Immunology, University of British Columbia

Bone marrow cells cultured with granulocyte macrophage colony stimulating factor (GM-CSF) generate a heterogeneous culture containing macrophages and dendritic cells (DCs). This method highlights using MHCII and hyaluronan (HA) binding to differentiate macrophages from the DCs in the GM-CSF culture. Macrophages in this culture have many similarities to alveolar macrophages.

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JoVE Journal

Experimental Procedure for Warm Spinning of Cast Aluminum Components
Matthew J. Roy 1, Daan M. Maijer 2
1School of Mechanical, Aerospace, and Civil Engineering, University of Manchester, 2Materials Engineering, University of British Columbia

An experimental protocol for instrumented warm rotary forming of cast aluminum alloys employing a bespoke industrially scaled apparatus is presented. Experimental considerations including thermal and mechanical effects are discussed, as well as similitude with full-scale processing of automotive wheels.

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Neuroscience

A Visual Guide to Sorting Electrophysiological Recordings Using 'SpikeSorter'
Nicholas V. Swindale 1, Catalin Mitelut 1, Timothy H. Murphy 2, Martin A. Spacek 1,3
1Ophthalmology and Visual Sciences, University of British Columbia, 2Department of Psychiatry, University of British Columbia, 3Neurobiology, Biology II, LMU München

The article shows how to use the program SpikeSorter to detect and sort spikes in extracellular recordings made with multi-electrode arrays.

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Immunology and Infection

System for Efficacy and Cytotoxicity Screening of Inhibitors Targeting Intracellular Mycobacterium tuberculosis
Xingji Zheng 1, Yossef Av-Gay 1
1Department of Medicine, University of British Columbia

We have developed a modular high-throughput screening system for discovering novel compounds against Mycobacterium tuberculosis, targeting intracellular and in-broth growing bacteria as well as cytotoxicity against the mammalian host cell.

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Bioengineering

Mammalian Cell Encapsulation in Alginate Beads Using a Simple Stirred Vessel
Corinne A. Hoesli 1, Roger L. J. Kiang 2, Kamini Raghuram 2, René G. Pedroza 3, Karen E. Markwick 1, Antonio M. R. Colantuoni 1, James M. Piret 2
1Department of Chemical Engineering, McGill University, 2Michael Smith Laboratories & Department of Chemical and Biological Engineering, University of British Columbia, 3Michael Smith Laboratories & Department of Pharmaceutical Sciences, University of British Columbia

This video and manuscript describe an emulsion-based method to encapsulate mammalian cells in 0.5% to 10% alginate beads which can be produced in large batches using a simple stirred vessel. The encapsulated cells can be cultured in vitro or transplanted for cellular therapy applications.

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Neuroscience

Determining Cell-surface Expression and Endocytic Rate of Proteins in Primary Astrocyte Cultures Using Biotinylation
Daniel Kai Long Tham 1, Hakima Moukhles 1
1Department of Cellular and Physiological Sciences, University of British Columbia

Two biotinylation-based methods, designed for determining the cell-surface expression and endocytic rate of proteins expressed at the plasma membrane, are presented in this report.

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Genetics

Generation of Native Chromatin Immunoprecipitation Sequencing Libraries for Nucleosome Density Analysis
Alireza Lorzadeh 1, Rodrigo Lopez Gutierrez 1, Linda Jackson 1, Michelle Moksa 1, Martin Hirst 1,2
1Department of Microbiology and Immunology, Michael Smith Laboratories Centre for High-Throughput Biology, University of British Columbia, 2Canada's Michael Smith Genome Science Center, BC Cancer Agency

We present a modified native chromatin immunoprecipitation sequencing (ChIP-seq) methodology for the generation of sequence datasets suitable for a nucleosome density ChIP-seq analytical framework integrating micrococcal nuclease (MNase) accessibility with histone modification measurements.

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Cancer Research

Practical Considerations in Studying Metastatic Lung Colonization in Osteosarcoma Using the Pulmonary Metastasis Assay
Michael M. Lizardo 1,2, Poul H. Sorensen 2,3
1Pediatric Oncology Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, 2BC Cancer Agency, Provincial Health Services Authority, 3Department of Pathology and Laboratory Medicine, University of British Columbia

The goal of this article is to provide a detailed description of the protocol for the pulmonary metastasis assay (PuMA). This model permits researchers to study metastatic osteosarcoma (OS) cell growth in lung tissue using a widefield fluorescence or confocal laser-scanning microscope.

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Immunology and Infection

Expression of Exogenous Antigens in the Mycobacterium bovis BCG Vaccine via Non-genetic Surface Decoration with the Avidin-biotin System
Ting-Yu Angela Liao 1, Alice Lau 1, Joseph Sunil 1, Vesa Hytönen 2, Zakaria Hmama 1
1Division of Infectious Diseases, Department of Medicine and Vancouver Costal Health Research Institute, University of British Columbia, 2Institute of Biomedical Technology, University of Tampere

A novel technique for rapid antigen display on a bacterial surface is presented, which involves surface biotinylation followed by exposure to proteins of interest in fusion with monomeric avidin. Loading BCG with selected antigens successfully improves its immunogenicity, suggesting that surface decoration can replace traditional genetic approaches.

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Immunology and Infection

Assessment of Antibody-based Drugs Effects on Murine Bone Marrow and Peritoneal Macrophage Activation
Lisa Kozicky 1, Laura M. Sly 1
1British Columbia Children's Hospital Research Institute, University of British Columbia

Antibody-based drugs have revolutionized treatment for inflammatory diseases. In addition to having direct effects on specific targets, antibodies can activate macrophages to become anti-inflammatory. This protocol describes how anti-inflammatory macrophage activation can be assessed in vitro, using mouse bone marrow macrophages, and in vivo, using peritoneal macrophages.

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Medicine

Routine Screening Method for Microparticles in Platelet Transfusions
Daniel Millar 1, Larry Murphy 2, Audrey Labrie 1, Elisabeth Maurer-Spurej 1,3,4
1Research & Development, LightIntegra Technology Inc., 2Quality Engineering & Regulatory, LightIntegra Technology Inc., 3Department of Pathology and Laboratory Medicine; Center for Blood Research, University of British Columbia, 4Canadian Blood Services

Platelet inventory management based on screening microparticle content in platelet concentrates is a new quality improvement initiative in hospital blood banks. The goal is to differentiate activated from non-activated platelets to optimize the use of platelets. Providing non-activated platelets to hematology-oncology patients might reduce their high risk to become refractory.

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Immunology and Infection

Visualizing the Actin and Microtubule Cytoskeletons at the B-cell Immune Synapse Using Stimulated Emission Depletion (STED) Microscopy
Jia C. Wang 1, Madison Bolger-Munro 1, Michael R. Gold 1
1Department of Microbiology and Immunology, University of British Columbia

We present a protocol for using STED microscopy to simultaneously image actin structures, microtubules, and microtubule plus-end binding proteins in B cells that have spread on coverslips coated with antibodies to the B-cell receptor, a model for the initial phase of immune synapse formation.

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Immunology and Infection

A Controlled Mouse Model for Neonatal Polymicrobial Sepsis
Byron Brook *1, Nelly Amenyogbe *1, Rym Ben-Othman 2, Bing Cai 2, Danny Harbeson 1, Freddy Francis 1, Aaron C. Liu 1, Natallia Varankovich 2, James Wynn 3,4, Tobias R. Kollmann 1,2
1Department of Experimental Medicine, University of British Columbia, 2Department of Pediatrics, Division of Infectious Diseases, University of British Columbia, 3Department of Pediatrics, College of Medicine, University of Florida, 4Department of Pathology, Immunology, and Laboratory Medicine, University of Florida

This protocol provides the necessary steps to establish and evaluate neonatal sepsis in 7-day-old mice.

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Immunology and Infection

Probiotic Studies in Neonatal Mice Using Gavage
Freddy Francis 1, Natallia Varankovich 2, Byron Brook 1, Nelly Amenyogbe 1, Rym Ben-Othman 2, Bing Cai 2, Danny Harbeson 1, Aaron C. Liu 1, Ben Dai 2, Shelly McErlane 4, Kris Andrews 4, Tobias R. Kollmann *1,2, Pinaki Panigrahi *3
1Department of Experimental Medicine, University of British Columbia, 2Department of Pediatrics, University of British Columbia, 3Department of Epidemiology and Pediatrics, University of Nebraska Medical Centre, 4Animal Care Services, University of British Columbia

This study details the process of gavaging precise amounts of probiotics to neonatal mice. The experimental set-up was optimized to include but is not limited to probiotic dosing, methods of administration, and quantification of bacteria in the intestines.

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Immunology and Infection

Chronic Salmonella Infection Induced Intestinal Fibrosis
Bernard C. Lo 1, Samuel B. Shin 1, Melina Messing 1, Kelly M. McNagny 1
1The Biomedical Research Centre, University of British Columbia

This protocol describes a mouse model of Salmonella driven intestinal fibrosis that resembles key pathological hallmarks of Crohn’s disease including transmural inflammation and fibrosis. This method can be used to evaluate host factors that alter fibrotic outcomes using mutant mice maintained on a C57Bl/6 genetic background.

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Biochemistry

PeptiQuick, a One-Step Incorporation of Membrane Proteins into Biotinylated Peptidiscs for Streamlined Protein Binding Assays
James W. Saville 1, Luca A. Troman 2, Franck Duong Van Hoa 1
1Department of Biochemistry, University of British Columbia, 2School of Biochemistry, University of Bristol

We present a method that combines membrane protein purification and reconstitution into peptidiscs in a single chromatographic step. Biotinylated scaffolds are used for direct surface attachment and measurement of protein-ligand interactions via biolayer interferometry.

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Immunology and Infection

Visualization of Gut Microbiota-host Interactions via Fluorescence In Situ Hybridization, Lectin Staining, and Imaging
Katharine M. Ng 1,2, Carolina Tropini 1,2,3
1School of Biomedical Engineering, University of British Columbia, 2Department of Microbiology and Immunology, University of British Columbia, 3Humans and the Microbiome Program, Canadian Institute for Advanced Research (CIFAR)

This streamlined protocol details a workflow to detect and image bacteria in complex tissue samples, from fixing the tissue to staining microbes with fluorescent in situ hybridization.

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JoVE Core

Capillary Electrophoresis-based Hydrogen/Deuterium Exchange for Conformational Characterization of Proteins with Top-down Mass Spectrometry
Lingxiao Chaihu 1,2, Xiaopeng Yao 1, Xiang Xu 1,2, Zhongqin Zhu 1, David D. Y. Chen 3, Guanbo Wang 1
1School of Chemistry and Materials Science, Nanjing Normal University, 2Institute for Cell Analysis, Shenzhen Bay Laboratory, 3Department of Chemistry, University of British Columbia

Presented here is a protocol for a capillary electrophoresis-based hydrogen/deuterium exchange (HDX) approach coupled with top-down mass spectrometry. This approach characterizes the difference in higher-order structures between different protein species, including proteins in different states and different proteoforms, by conducting concurrent differential HDX and electrophoretic separation.

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Environment

Enhanced Reproducibility and Precision of High-Throughput Quantification of Bacterial Growth Data Using a Microplate Reader
Leili Abkar 1, Florentin M. Wilfart 2, Marta Piercey 3, Graham A. Gagnon 4
1Chemical and Biological Engineering Department, University of British Columbia, 2Process Engineering and Applied Science, Dalhousie University, 3Food Science and Technology, Dalhousie University, 4Center for Water Resources Studies, Civil and Resource Engineering, Dalhousie University

Here, a high-throughput protocol is presented to measure growth data, including growth curves, growth rate, and maximum growth rate. The protocol was verified and validated using two biofilm-producing bacteria. The results and approach applied in this study can be expanded to other high-throughput protocols using microplate readers.

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Neuroscience

Assessing Changes in Synaptic Plasticity Using an Awake Closed-Head Injury Model of Mild Traumatic Brain Injury
Brian R. Christie 1,2, Allyson Gross 1, Annika Willoughby 1, Erin Grafe 1, Justin Brand 1, Emily Bosdachin 1, Hannah M. O. Reid 1, Crystal Acosta 1, Eric Eyolfson 1
1Division of Medical Sciences, University of Victoria, 2Island Medical Program, University of British Columbia

Here, it is demonstrated how an awake closed-head injury model can be used for examining the effects of repeated mild traumatic brain injury (r-mTBI) on synaptic plasticity in the hippocampus. The model replicates important features of r-mTBI in patients and is used in conjunction with in vitro electrophysiology.

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Bioengineering

Generation of Human Blood Vessel Organoids from Pluripotent Stem Cells
Nicolas Werschler 1,2, Josef Penninger 1,2,3,4
1School of Biomedical Engineering, University of British Columbia, 2Life Sciences Institute, University of British Columbia, 3Department of Medical Genetics, University of British Columbia, 4Institute of Molecular Biotechnology of the Austrian Academy of Sciences (IMBA)

This protocol describes the generation of self-organizing blood vessels from human pluripotent and induced pluripotent stem cells. These blood vessel networks exhibit an extensive and connected endothelial network surrounded by pericytes, smooth muscle actin, and a continuous basement membrane.

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Bioengineering

Rapid Development of Cell State Identification Circuits with Poly-Transfection
Noreen Wauford 1, Ross Jones 1,2, Charles Van De Mark 3, Ron Weiss 1,3
1Department of Biological Engineering, Massachusetts Institute of Technology, 2School of Biological Engineering, University of British Columbia, 3Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology

Complex genetic circuits are time-consuming to design, test, and optimize. To facilitate this process, mammalian cells are transfected in a way that allows the testing of multiple stoichiometries of circuit components in a single well. This protocol outlines the steps for experimental planning, transfection, and data analysis.

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Developmental Biology

Differentiation of Human Pluripotent Stem Cells into Insulin-Producing Islet Clusters
Jia Zhao *1, Shenghui Liang *1, Mitchell J. S. Braam 1, Robert K. Baker 1, Diepiriye G. Iworima 1,2, Nina Quiskamp 3, Timothy J. Kieffer 1,2,4
1Department of Cellular & Physiological Sciences, Life Science Institute, University of British Columbia, 2School of Biomedical Engineering, University of British Columbia, 3STEMCELL Technologies Inc, 4Department of Surgery, University of British Columbia

The differentiation of stem cells into islet cells provides an alternative solution to conventional diabetes treatment and disease modeling. We describe a detailed stem cell culture protocol that combines a commercial differentiation kit with a previously validated method to aid in producing insulin-secreting, stem cell-derived islets in a dish.

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Neuroscience

Quantification of Global Histone Post Translational Modifications Using Intranuclear Flow Cytometry in Isolated Mouse Brain Microglia
Morgan Towriss 1, Jennifer Kim 2, Annie Vogel Ciernia 1
1Department of Biochemistry and Molecular Biology, Djavad Mowafaghian Centre for Brain Health, University of British Columbia, 2Graduate Program in Neuroscience, Djavad Mowafaghian Centre for Brain Health, University of British Columbia

This work describes a protocol for the quantification of global histone modifications using intranuclear flow cytometry in isolated brain microglia. The work also contains the microglia isolation protocol that was used for data collection.

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Medicine

Measuring Maxillary Posterior Tooth Movement: A Model Assessment using Palatal and Dental Superimposition
Abdulraheem A. Alwafi 1,2, Scott Panther 2, Angela Lo 2, Edwin H. Yen 2, Bingshuang Zou 2
1Department of Dental Public Health, Faculty of Dentistry, King Abdulaziz University, 2Department of Oral Health Science, Faculty of Dentistry, University of British Columbia

This manuscript presents a comprehensive protocol to evaluate the three-dimensional (3D) movement of maxillary posterior teeth with clear aligners using digital model superimposition, an invaluable tool in orthodontics and dentofacial orthopedics.

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Developmental Biology

An Optimized Mouse Embryonic Stem Cell Based Reverse Poly-Transfection Technique for Rapid Exploration of Nucleic Acid Ratios
Kieran Maheden 1, Karen Hwang 1, Ipek Egilmez 1, Nika Shakiba 1
1School of Biomedical Engineering, University of British Columbia

The present protocol describes a method for reverse poly-transfection of mouse embryonic stem cells during culture with 2i and LIF media. This method yields higher viability and efficiency than traditional forward transfection protocols, while also enabling one-pot optimization of plasmid ratios.

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Immunology and Infection

T4 Bacteriophage and E. coli Interaction in the Murine Intestine: A Prototypical Model for Studying Host-Bacteriophage Dynamics In Vivo
Nicola Pett 1, Michael Hunter 1, Natalia A. Carranza García 1, Jung Hee Seo 1, Samuel R. Collins 1, Forest Rohwer 2, Lisa C. Osborne 1, Carolina Tropini 1,3,4
1Department of Microbiology and Immunology, Life Sciences Institute, University of British Columbia, 2Department of Biology, San Diego State University, 3School of Biomedical Engineering, University of British Columbia, 4Humans and the Microbiome Program, Canadian Institute for Advanced Research

Bacteriophages (phages), viruses that infect bacteria, are an integral component of the gut microbiome. Though these symbiotic inhabitants drive bacterial fitness and population dynamics, little is understood about how they impact gut homeostasis and disease. This protocol studies isolated T4 phages within a mouse model, adaptable to other phage-bacterial pairs.

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Medicine

Reliability of Artificial Intelligence-Based Cone Beam Computed Tomography Integration with Digital Dental Images
Ju-Heon Lee *1, Nam-Ki Lee *1, Bingshuang Zou 2, Jae Hyun Park 3,4, Tae-Hyun Choi 1
1Department of Orthodontics, Seoul National University Bundang Hospital, 2Division of Orthodontics, Faculty of Dentistry, University of British Columbia, 3Postgraduate Orthodontic Program, Arizona School of Dentistry & Oral Health, A. T. Still University, 4Graduate School of Dentistry, Kyung Hee University

A process of registering cone-beam computed tomography scans and digital dental images has been presented using artificial intelligence (AI) -assisted identification of landmarks and merging. A comparison with surface-based registration shows that AI-based digitization and integration are reliable and reproducible.

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Medicine

Multi-modal Pulmonary Imaging: Using Complementary Information from CT and Hyperpolarized 129Xe MRI to Evaluate Lung Structure-Function
Rachel L Eddy 1,2, George H Xu 3, Jonathon A Leipsic 1,4, Janice M Leung 1,2, Don D Sin 1,2, Chase S Hall 5, Roger C Tam 3,4
1UBC Centre for Heart Lung Innovation, St. Paul’s Hospital, 2Division of Respiratory Medicine, Department of Medicine, University of British Columbia, 3School of Biomedical Engineering, University of British Columbia, 4Department of Radiology, University of British Columbia, 5Division of Pulmonary and Critical Care Medicine, University of Kansas Medical Center

CT and 129Xe MRI provide complementary lung structure-function information that can be exploited for regional analysis using image registration. Here, we provide a protocol that builds from the existing literature for 129Xe MR to CT image registration using open-source platforms.

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Fibro-Adipogenic Progenitor Isolation, Expansion, and Differentiation from the Spiny Mouse Model
Bruce Lin 1, Hesham Soliman 1,2, Fabio M. V. Rossi 1, Marine Theret 1
1School of Biomedical Engineering and Department of Medical Genetics, University of British Columbia, 2Aspect Biosystems

This protocol outlines the isolation of skeletal and cardiac muscle fibro-adipogenic progenitors (FAPs) from spiny mouse (Acomys) via enzymatic dissociation and fluorescence-activated cell sorting. The FAPs obtained from this protocol can be effectively expanded and differentiated to myofibroblasts and adipocytes.

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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 Bottiglieri 4, 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.

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Environment

Measurements of Soil Water Potential and Conductivity based on a Simple Evaporation Experiment using a Hydraulic Property Analyzer
Alessia J. Marchesan 1, Kris Guenette 1, Lewis K. Fausak 2, Guillermo Hernandez Ramirez 1
1Renewable Resources, University of Alberta, 2Faculty of Land and Food Systems, University of British Columbia

This article features a simple evaporation experiment using a hydraulic property instrument for a soil sample. Through efficient means, measurements can be taken over a series of days to generate high-quality data.

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Neuroscience

Implantation of Optoelectronic Devices in the Rodent Spinal Cord
Shahriar Shalileh *1,2, Adan Moallemi *1,2, Brianna Tsuyuki 1,2, Aiden A. P. Simard 2, Dena Shahriari 1,2,3,4
1School of Biomedical Engineering, University of British Columbia, 2International Collaborations on Repair Discoveries (ICORD), 3Department of Orthopaedics, University of British Columbia, 4Institute for Computing, Information and Cognitive Systems, University of British Columbia

This protocol details a surgical procedure for performing spinal cord surgery and for implanting and securing an optical shank over the spinal cord in rodents.

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