University of Ottawa View Institution's Website 41 articles published in JoVE Bioengineering Decellularized Apple-Derived Scaffolds for Bone Tissue Engineering In Vitro and In Vivo Maxime Leblanc Latour1, Maryam Tarar2, Ryan J. Hickey1, Charles M. Cuerrier1, Isabelle Catelas3,4,5, Andrew E. Pelling1,2,6,7 1Department of Physics, University of Ottawa, 2Department of Biology, University of Ottawa, 3Department of Mechanical Engineering, University of Ottawa, 4Department of Surgery, University of Ottawa, 5Department of Biochemistry, Microbiology and Immunology, University of Ottawa, 6Institute for Science, Society and Policy, University of Ottawa, 7SymbioticA, School of Human Sciences, University of Western Australia In this study, we detail methods of decellularization, physical characterization, imaging, and in vivo implantation of plant-based biomaterials, as well as methods for cell seeding and differentiation in the scaffolds. The described methods allow the evaluation of plant-based biomaterials for bone tissue engineering applications. Biochemistry ABCG5/G8 Crystallization in a Lipidic Bicelle Environment for X-Ray Crystallography Sarah Wazir1, Danny Farhat1, Mahalashmi Srinivasan1,2, Jyh-Yeuan Lee1 1Department of Biochemistry, Microbiology and Immunology, Faculty of Medicine, University of Ottawa, 2Department of Biochemistry, Faculty of Medicine and Dentistry, University of Alberta This protocol describes a setup for the crystallization of the sterol transporter ABCG5/G8. ABCG5/G8 is reconstituted into bicelles for hanging-drop crystallization. The protocol does not require specialized materials or substrates, making it accessible and easy to adapt in any laboratory for determining the protein structure through X-ray crystallography. Biology Viral Transgene Expression in Rodent Hearts and the Assessment of Cardiac Arrhythmia Risk Aizhu Lu1,2, Jerry Wang1,2, Ying Xia1,2, Ruonan Gu1, Kyoung-Han Kim1,2, Erin E. Mulvihill1,3, Darryl R. Davis1,2, Rob S. Beanlands1, Wenbin Liang1,2 1University of Ottawa Heart Institute, 2Department of Cellular and Molecular Medicine, University of Ottawa, 3Department of Biochemistry, Microbiology and Immunology, University of Ottawa The present protocol describes methods for transgene expression in rat and mouse hearts by direct intramyocardial injection of the virus under echocardiography guidance. Methods for the assessment of the susceptibility of hearts to ventricular arrhythmias by the programmed electrical stimulation of isolated, Langendorff-perfused hearts are also explained here. Biochemistry Transverse Sectioning of Mature Rice (Oryza sativa L.) Kernels for Scanning Electron Microscopy Imaging Using Pipette Tips as Immobilization Support Jordan Demone1, Kiah A. Barton1,2, Illimar Altosaar1,2 1Department of Biochemistry, Microbiology, and Immunology, University of Ottawa, 2Proteins Easy Corp This protocol allows for the preparation of transverse sections of cereal seeds (e.g., rice) for the analysis of endosperm and starch granule morphology using scanning electron microscopy. Immunology and Infection Detection of SARS-CoV-2 Receptor-Binding Domain Antibody using a HiBiT-Based Bioreporter Reza Rezaei1,2, Abera Surendran1,2, Ragunath Singaravelu1,2, Taylor R. Jamieson1,2, Parisa Taklifi3, Joanna Poutou1,2, Taha Azad1,2, Carolina S. Ilkow1,2 1Ottawa Hospital Research Institute, 2Department of Biochemistry, Microbiology and Immunology, University of Ottawa, 3Department of Biotechnology, University of Tehran The outlined protocol describes the procedure for producing the HiBiT-receptor-binding domain protein complex and its application for fast and sensitive detection of SARS-CoV-2 antibodies. Immunology and Infection Detection of SARS-CoV-2 Neutralizing Antibodies using High-Throughput Fluorescent Imaging of Pseudovirus Infection Taylor R. Jamieson1,2, Joanna Poutou1,2, Ricardo Marius1,2, Xiaohong He1, Reza Rezaei1,2, Taha Azad1,2, Carolina S. Ilkow1,2 1Ottawa Hospital Research Institute, 2Department of Biochemistry, Microbiology and Immunology, University of Ottawa The protocol described here outlines a fast and effective method for measuring neutralizing antibodies against the SARS-CoV-2 spike protein by evaluating the ability of convalescent serum samples to inhibit infection by an enhanced green fluorescent protein-labeled vesicular stomatitis virus pseudotyped with spike glycoprotein. Chemistry Hyperspectral Imaging as a Tool to Study Optical Anisotropy in Lanthanide-Based Molecular Single Crystals Emille M. Rodrigues1, Nelson Rutajoga1, David Rioux2, Jacob Yvon-Leroux2, Eva Hemmer1 1Department of Chemistry and Biomolecular Sciences, University of Ottawa, 2Photon etc Here, we present a protocol to obtain luminescent hyperspectral imaging data and to analyze optical anisotropy features of lanthanide-based single crystals using a Hyperspectral Imaging System. Biochemistry Parallel Interrogation of β-Arrestin2 Recruitment for Ligand Screening on a GPCR-Wide Scale using PRESTO-Tango Assay Manel Zeghal1, Geneviève Laroche1, Patrick M. Giguère1,2 1Department of Biochemistry, Microbiology and Immunology, University of Ottawa, 2Brain and Mind Research Institute, University of Ottawa Given that GPCRs are attractive druggable targets, GPCR ligand screening is thus indispensable for the identification of lead compounds and for deorphanization studies. Towards these efforts, we describe PRESTO-Tango, an open-source resource platform used for simultaneous profiling of transient β-arrestin2 recruitment at approximately 300 GPCRs using a TEV-based reporter assay. Behavior Examining Bilingual Language Control Using the Stroop Task Laura Sabourin*1, Santa Vīnerte*1 1Department of Linguistics, University of Ottawa This bilingual Stroop task uses Congruent, Incongruent, and Neutral stimuli presented in blocks in the first language (L1) only, the second language (L2) only, and a combination of L1 and L2. This task allows for an examination of language processing and cognitive control in both L1 and L2. Biology Assessment of the Metabolic Effects of Isocaloric 2:1 Intermittent Fasting in Mice Ri Youn Kim*1,2, Ju Hee Lee*3,4, Yena Oh1,2, Hoon-Ki Sung3,4,5, Kyoung-Han Kim1,2 1University of Ottawa Heart Institute, 2Department of Cellular and Molecular Medicine, University of Ottawa, 3Translational Medicine Program, The Hospital for Sick Children, 4Department of Laboratory Medicine and Pathobiology, University of Toronto, 5Banting and Best Diabetes Centre, University of Toronto The current article describes a detailed protocol for isocaloric 2:1 intermittent fasting to protect and treat against obesity and impaired glucose metabolism in wild-type and ob/ob mice. Engineering Fabricating van der Waals Heterostructures with Precise Rotational Alignment Justin Boddison-Chouinard1, Ryan Plumadore1, Adina Luican-Mayer1 1Department of Physics, University of Ottawa In this work we describe a technique that is used to create new crystals (van der Waals heterostructures) by stacking ultrathin layered 2D materials with precise control over position and relative orientation. Biochemistry Hybrid Clear/Blue Native Electrophoresis for the Separation and Analysis of Mitochondrial Respiratory Chain Supercomplexes Alexanne Cuillerier1, Yan Burelle1 1Interdisciplinary School of Health Science, Faculty of Health Sciences and Department of Cellular and Molecular Medicine, Faculty of Medicine, University of Ottawa Here we present a protocol to extract, resolve and identify mitochondrial supercomplexes which minimizes exposure to detergents and Coomassie Blue. This protocol offers an optimal balance between resolution, and preservation of enzyme activities, while minimizing the risk of losing labile protein-protein interactions. Environment An Anaerobic Biosensor Assay for the Detection of Mercury and Cadmium Benjamin R. Stenzler1, Jessica Gaudet1, Alexandre J. Poulain1 1Department of Biology, University of Ottawa Here, we present a protocol to use an anaerobic whole-cell microbial biosensor to evaluate how different environmental variables affect the bioavailability of Hg and Cd to bacteria in anoxic environments. Medicine Using a Knee Arthrometer to Evaluate Tissue-specific Contributions to Knee Flexion Contracture in the Rat T. Mark Campbell1,2,3, Katherine Reilly2,3, Louis Goudreau4, Odette Laneuville2,5, Hans Uhthoff2,3, Guy Trudel2,3,4 1Elizabeth Bruyère Hospital, 2The Bone and Joint Research Laboratory, University of Ottawa, 3Department of Medicine, University of Ottawa, 4The Ottawa Hospital Rehabilitation Centre, 5Department of Biology, University of Ottawa The goal of the protocol is to measure the extension range of motion of the rat knee. The effects of various diseases that increase the stiffness of the knee joint and the effectiveness of treatments can be quantified. Genetics High-throughput DNA Extraction and Genotyping of 3dpf Zebrafish Larvae by Fin Clipping Ceres Kosuta1,2, Kate Daniel1, Devon L. Johnstone1,2, Kevin Mongeon1,2, Kevin Ban1,2, Sophie LeBlanc1, Stuart MacLeod1, Karim Et-Tahiry2, Marc Ekker2, Alex MacKenzie1, Izabella Pena1,2 1 Zebrafish have been used as reliable genetic model organisms in biomedical research, especially with the advent of gene-editing technologies. When larval phenotypes are expected, DNA extraction and genotype identification can be challenging. Here, we describe an efficient genotyping procedure for zebrafish larvae, by tail clipping, as early as 72-h post-fertilization. Genetics Targeted Next-generation Sequencing and Bioinformatics Pipeline to Evaluate Genetic Determinants of Constitutional Disease Allison A. Dilliott1,2, Sali M.K. Farhan3, Mahdi Ghani4, Christine Sato4, Eric Liang5, Ming Zhang4, Adam D. McIntyre1, Henian Cao1, Lemuel Racacho6,7, John F. Robinson1, Michael J. Strong1,8, Mario Masellis9,10, Dennis E. Bulman6,7, Ekaterina Rogaeva4, Anthony Lang10,11, Carmela Tartaglia4,10, Elizabeth Finger12,13, Lorne Zinman9, John Turnbull14, Morris Freedman10,15, Rick Swartz9, Sandra E. Black9,16, Robert A. Hegele1,2 1Robarts Research Institute, Schulich School of Medicine and Dentistry, Western University, 2Department of Biochemistry, Schulich School of Medicine and Dentistry, Western University, 3Analytic and Translational Genetics Unit, Center for Genomic Medicine, Harvard Medical School, Massachusetts General Hospital, Stanley Centre for Psychiatric Research, Broad Institute of MIT and Harvard, 4Tanz Centre for Research in Neurodegenerative Diseases, University of Toronto, 5 Targeted next-generation sequencing is a time- and cost-efficient approach that is becoming increasingly popular in both disease research and clinical diagnostics. The protocol described here presents the complex workflow required for sequencing and the bioinformatics process used to identify genetic variants that contribute to disease. Cancer Research Genome-wide RNAi Screening to Identify Host Factors That Modulate Oncolytic Virus Therapy Kristina J. Allan1,2, Douglas J. Mahoney1,4, Stephen D. Baird1, Charles A. Lefebvre1, David F. Stojdl1,2,3 1 Here we describe a protocol for employing high-throughput RNAi screening to uncover host targets that can be manipulated to enhance oncolytic virus therapy, specifically rhabodvirus and vaccinia virus therapy, but it can be readily adapted to other oncolytic virus platforms or for discovering host genes that modulate virus replication generally. Engineering All-electronic Nanosecond-resolved Scanning Tunneling Microscopy: Facilitating the Investigation of Single Dopant Charge Dynamics Mohammad Rashidi1,2, Wyatt Vine1, Jacob A.J. Burgess3,4,5, Marco Taucer1,2,6, Roshan Achal1, Jason L. Pitters2, Sebastian Loth3,4, Robert A. Wolkow1,2 1Department of Physics, University of Alberta, 2National Institute for Nanotechnology, National Research Council of Canada, Edmonton, 3Max Planck Institute for the Structure and Dynamics of Matter, 4Max Planck Institute for Solid State Research, 5Department of Physics and Astronomy, University of Manitoba, 6Joint Attosecond Science Laboratory, University of Ottawa We demonstrate an all-electronic method to observe nanosecond-resolved charge dynamics of dopant atoms in silicon with a scanning tunneling microscope. Neuroscience Modeling Neuronal Death and Degeneration in Mouse Primary Cerebellar Granule Neurons Matthew Laaper1,2, Takrima Haque1, Ruth S. Slack3, Arezu Jahani-Asl1,2,4 1Lady Davis Institute for Medical Research, Jewish General Hospital, 2Integrated Program in Neuroscience, McGill University, 3Department of Cellular and Molecular Medicine, University of Ottawa, 4Department of Oncology, Faculty of Medicine, McGill University This protocol describes a simple method for isolating and culturing primary mouse cerebral granule neurons (CGNs) from 6-7 day old pups, efficient transduction of CGNs for loss and gain of function studies, and modelling NMDA-induced neuronal excitotoxicity, low-potassium-induced cell death, DNA-damage, and oxidative stress using the same culture model. Genetics Inducible T7 RNA Polymerase-mediated Multigene Expression System, pMGX Mohamed I. Hassan*1, Fern R. McSorley*1, Kinya Hotta2, Christopher N. Boddy1 1Department of Chemistry and Biomolecular Sciences, University of Ottawa, 2School of Biosciences, The University of Nottingham Malaysia Campus This study describes methods for the T7-mediated co-expression of multiple genes from a single plasmid in Escherichia coli using the pMGX plasmid system. Biology Isolation of CD146+ Resident Lung Mesenchymal Stromal Cells from Rat Lungs Jennifer J.P. Collins1,2, Marius A. Möbius1,3,4, Bernard Thébaud1,2,5 1Sinclair Centre for Regenerative Medicine, Ottawa Hospital Research Institute, 2University of Ottawa, 3Department of Neonatology and Pediatric Critical Care Medicine, Medical Faculty and University Hospital Carl Gustav Carus, Technische Universität Dresden, 4DFG Research Center and Cluster of Excellence for Regenerative Therapies (CRTD), Technische Universität, Dresden, 5 This protocol describes an isolation technique for obtaining primary lung resident mesenchymal stromal cells from rats, through the use of enzymatic digestion, density gradient separation, plastic adherence and CD146+ magnetic bead selection. Behavior Substantiating Appropriate Motion Capture Techniques for the Assessment of Nordic Walking Gait and Posture in Older Adults Christopher M. Dalton1, Julie Nantel1 1School of Human Kinetics, Faculty of Health Sciences, University of Ottawa The aim was to substantiate optimal use of data collection techniques for Nordic walking gait and posture analysis. Three-dimensional motion capture should be used during short duration analysis (i.e. single gait cycle), while accelerometry should be employed for longer duration analysis (i.e. repeated cycles) like a 6 Minute Walk Test. Behavior Evaluation of a Smartphone-based Human Activity Recognition System in a Daily Living Environment Edward D. Lemaire1, Marco D. Tundo2, Natalie Baddour2 1The Ottawa Hospital Research Institute, University of Ottawa, 2Department of Mechanical Engineering, University of Ottawa A standardized evaluation method was developed for Wearable Mobility Monitoring Systems (WMMS) that includes continuous activities in a realistic daily living environment. Testing with a series of daily living activities can decrease activity recognition sensitivity; therefore, realistic testing circuits are encouraged for valid evaluation of WMMS performance. 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. Developmental Biology A Time Differential Staining Technique Coupled with Full Bilateral Gill Denervation to Study Ionocytes in Fish Velislava Tzaneva1, Steve F. Perry1 1Department of Biology, University of Ottawa This manuscript describes a protocol to track the re-distribution of branchial ionocytes and their innervation using a time differential staining technique coupled with full bilateral gill denervation. Developmental Biology Induction of Protein Deletion Through In Utero Electroporation to Define Deficits in Neuronal Migration in Transgenic Models Devon S. Svoboda1, Alysen Clark1, David S. Park1, Ruth S. Slack1 1Department of Cellular and Molecular Medicine, University of Ottawa Neuron migration is regulated by numerous cell autonomous and non-cell autonomous factors. This protocol shows how in utero electroporation can be used to determine whether a phenotype in a transgenic mouse is due to disruption of a cell intrinsic mechanism or impairment of interaction between the neuron and its environment. Neuroscience Radio Frequency Identification and Motion-sensitive Video Efficiently Automate Recording of Unrewarded Choice Behavior by Bumblebees Levente L. Orbán1, Catherine M.S. Plowright1 1School of Psychology, University of Ottawa This video describes Radio-Frequency Identification (RFID) and motion-sensitive video recording methods to monitor choice behavior by bumblebees. Biology Assessment of Selective mRNA Translation in Mammalian Cells by Polysome Profiling Mame Daro Faye1, Tyson E Graber2, Martin Holcik3 1Apoptosis Research Centre, Children's Hospital of Eastern Ontario Research Institute and Department of Biochemistry, Microbiology and Immunology, University of Ottawa, 2Montreal Neurological Institute, 3Apoptosis Research Centre, Children's Hospital of Eastern Ontario Research Institute and Department of Pediatrics, University of Ottawa The ability of cells to adapt to stress is crucial for their survival. Regulation of mRNA translation is one such adaptation strategy, providing for rapid regulation of the proteome. Here, we provide a standardized polysome profiling protocol to identify specific mRNAs that are selectively translated under stress conditions. Biology High-throughput Titration of Luciferase-expressing Recombinant Viruses Vanessa Garcia1,2, Ramya Krishnan1,2, Colin Davis1,2, Cory Batenchuk1,2, Fabrice Le Boeuf1,3, Hesham Abdelbary1,3, Jean-Simon Diallo1,2 1Center for Innovative Cancer Research, Ottawa Hospital Research Institute, 2Department of Biochemistry, Microbiology and Immunology, Faculty of Medicine, University of Ottawa, 3Faculty of Medicine, University of Ottawa This article presents a high-throughput luciferase expression-based method of titrating various RNA and DNA viruses using automated and manual liquid handlers. Bioengineering A Novel Stretching Platform for Applications in Cell and Tissue Mechanobiology Dominique Tremblay1, Charles M. Cuerrier1,2, Lukasz Andrzejewski1, Edward R. O'Brien3, Andrew E. Pelling1,4,5 1Centre for Interdisciplinary NanoPhysics, Department of Physics, University of Ottawa, 2University of Ottawa Heart Institue, University of Ottawa, 3Libin Cardiovascular Institute of Alberta, University of Calgary, 4Department of Biology, University of Ottawa, 5Institute for Science, Society and Policy, University of Ottawa We present in this article a novel stretching platform that can be used to investigate single cell responses to complex anisotropic biaxial mechanical deformation and quantify the mechanical properties of biological tissues. Medicine A Mouse Tumor Model of Surgical Stress to Explore the Mechanisms of Postoperative Immunosuppression and Evaluate Novel Perioperative Immunotherapies Lee-Hwa Tai1, Christiano Tanese de Souza1, Shalini Sahi1, Jiqing Zhang1,2,4, Almohanad A Alkayyal1,3,5, Abhirami Anu Ananth1,3, Rebecca A.C. Auer1,6 1Centre for Innovative Cancer Research, Ottawa Hospital Research Institute, 2Department of Cellular and Molecular Medicine, University of Ottawa, 3Department of Biochemistry, Microbiology and Immunology, University of Ottawa, 4Department of Neurosurgery, The Second Hospital of Shandong University, 5Department of Medical Laboratory Technology, University of Tabuk, 6Department of Surgery, Ottawa General Hospital A mouse tumor model of surgical stress is used to explore how postoperative immune suppression promotes metastatic disease and to evaluate immunostimulatory perioperative therapies. Neuroscience Long-term Behavioral Tracking of Freely Swimming Weakly Electric Fish James J. Jun1,2,3, André Longtin1,2,3, Leonard Maler2,3 1Department of Physics, University of Ottawa, 2Department of Cellular and Molecular Medicine, University of Ottawa, 3Centre for Neural Dynamics, University of Ottawa We describe a set of techniques for studying spontaneous behavior of freely swimming weakly electric fish over an extended period of time, by synchronously measuring the animal's electric organ discharge timing, body position and posture both accurately and reliably in a specially designed aquarium tank inside a sensory isolation chamber. Immunology and Infection Establishing the Minimal Bactericidal Concentration of an Antimicrobial Agent for Planktonic Cells (MBC-P) and Biofilm Cells (MBC-B) Thien-Fah Mah1 1Department of Biochemistry, Microbiology and Immunology, University of Ottawa This protocol allows for a direct comparison between planktonic and biofilm resistance for a bacterial strain that can form a biofilm in vitro using a 96-well microtiter plate. Planktonic or biofilm bacteria are exposed to serial dilutions of the antimicrobial agent of choice. Viability is assayed by growth on agar plates. Engineering Fine-tuning the Size and Minimizing the Noise of Solid-state Nanopores Eric Beamish1, Harold Kwok1, Vincent Tabard-Cossa1, Michel Godin1,2 1Department of Physics, University of Ottawa, 2Ottawa-Carleton Institute of Biomedical Engineering, University of Ottawa A methodology for preparing solid-state nanopores in solution for biomolecular translocation experiments is presented. By applying short pulses of high electric fields, the nanopore diameter can be controllably enlarged with subnanometer precision and its electrical noise characteristics significantly improved. This procedure is performed in situ using standard laboratory equipment under experimental conditions. Bioengineering Micro-particle Image Velocimetry for Velocity Profile Measurements of Micro Blood Flows Katie L. Pitts1, Marianne Fenech1,2 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. Biology Isolation and Culture of Individual Myofibers and their Satellite Cells from Adult Skeletal Muscle Alessandra Pasut1,2, Andrew E. Jones1,2, Michael A. Rudnicki1,2 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. Biology Performing Vaginal Lavage, Crystal Violet Staining, and Vaginal Cytological Evaluation for Mouse Estrous Cycle Staging Identification Ashleigh C. McLean1,2,3, Nicolas Valenzuela3,4, Stephen Fai3,4, Steffany A.L. Bennett1,3 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. Neuroscience Derivation of Enriched Oligodendrocyte Cultures and Oligodendrocyte/Neuron Myelinating Co-cultures from Post-natal Murine Tissues Ryan W. O'Meara1,2, Scott D. Ryan1, Holly Colognato3, Rashmi Kothary1,2,4 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 Lentiviral-mediated Knockdown During Ex Vivo Erythropoiesis of Human Hematopoietic Stem Cells Carmen G. Palii1, Roya Pasha1, Marjorie Brand1,2 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. Immunology and Infection Quantitative Analyses of all Influenza Type A Viral Hemagglutinins and Neuraminidases using Universal Antibodies in Simple Slot Blot Assays Caroline Gravel*1, Changgui Li*2, Junzhi Wang2, Anwar M Hashem1,3,4, Bozena Jaentschke1, Gary Van Domselaar5, Runtao He5, Xuguang Li1,3 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. 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.