National Research Council Canada View Institution's Website 10 articles published in JoVE Immunology and Infection Generation of Null Mutants to Elucidate the Role of Bacterial Glycosyltransferases in Bacterial Motility Juan M. Tomás1, Kelly M. Fulton2,3, Susan M. Twine2,4, Susana Merino1 1Departamento de Genética, Microbiología y Estadística, Universidad de Barcelona, 2Human Health Therapeutics Research Centre, National Research Council Canada, 3Department of Chemistry, Carleton University, 4Department of Biology, Carleton University Here, we describe the construction of null mutants of Aeromonas in specific glycosyltransferases or regions containing glycosyltransferases, the motility assays, and flagella purification performed to establish the involvement and function of their encoded enzymes in the biosynthesis of a glycan, as well as the role of this glycan in bacterial pathogenesis. Bioengineering Fabrication of a Crystalline Nanocellulose Embedded Agarose Biomaterial Ink for Bone Marrow-Derived Mast Cell Culture Leshern Karamchand1, Ashley Wagner1, Syed Benazir Alam1, Marianna Kulka1,2 1Nanotechnology Research Center, National Research Council Canada, 2Department of Medical Microbiology and Immunology, University of Alberta This protocol highlights a method to rapidly assess the biocompatibility of a crystalline nanocellulose (CNC)/agarose composite hydrogel biomaterial ink with mouse bone marrow-derived mast cells in terms of cell viability and phenotypic expression of the cell surface receptors, Kit (CD117) and high-affinity IgE receptor (FcεRI). Bioengineering Methylation Specific Multiplex Droplet PCR using Polymer Droplet Generator Device for Hematological Diagnostics Lidija Malic1, Abdelrahman Elmanzalawy1, Jamal Daoud2, Matthias Geissler1, Alex Boutin1, Ljuboje Lukic1, Mojra Janta1, Dillon Da Fonte1, Christina Nassif1, Teodor Veres1 1Life Sciences Division, National Research Council of Canada, 2Galenvs Sciences, Inc. Epigenetic markers are used for white blood cell (WBC) subtyping through the quantification of DNA methylation patterns. This protocol presents a multiplex droplet polymerase chain reaction (mdPCR) method using a thermoplastic elastomer (TPE)-based microfluidic device for droplet generation allowing for precise and multiplex methylation-specific target quantification of WBC differential counts. Engineering Intermediate Strain Rate Material Characterization with Digital Image Correlation Meysam Rahmat1, David Backman1, Richard Desnoyers1 1Aerospace Research Centre, National Research Council Canada Here we present a methodology for the dynamic characterization of tensile specimens at intermediate strain rates using a high-speed servo-hydraulic load frame. Procedures for strain gauge instrumentation and analysis, as well as for digital image correlation strain measurements on the specimens, are also defined. 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. Bioengineering Engineering 3D Cellularized Collagen Gels for Vascular Tissue Regeneration Sébastien Meghezi1, Dawit G. Seifu1,2, Nina Bono1,3, Larry Unsworth2,4,5, Kibret Mequanint6, Diego Mantovani1,2 1Laboratory for Biomaterials and Bioengineering, Department Min-Met-Materials Eng & CHU de Québec Research Center, Canada Research Chair I for the Innovation in Surgery, Laval University, 2NSERC CREATE Program for Regenerative Medicine (NCPRM), Laval University, 3Department Electronics, Information and Bioengineering, Politecnico di Milano, 4Department of Chemical and Materials Engineering, University of Alberta, 5National Institute for Nanotechnology, National Research Council (Canada), 6Department of Chemical and Biochemical Engineering, University of Western Ontario In this work, we present a technique for the rapid fabrication of living vascular tissues by direct culturing of collagen, smooth muscle cells and endothelial cells. In addition, a new protocol for the mechanical characterization of engineered vascular tissues is described. Engineering Surface Enhanced Raman Spectroscopy Detection of Biomolecules Using EBL Fabricated Nanostructured Substrates Robert F. Peters1,2, Luis Gutierrez-Rivera1,2, Steven K. Dew1,2, Maria Stepanova1,2 1Department of Electrical and Computer Engineering, University of Alberta, 2National Institute for Nanotechnology, National Research Council of Canada We describe the fabrication and characterization of nano-biological systems interfacing nanostructured substrates with immobilized proteins and aptamers. The relevant experimental steps involving lithographic fabrication of nanostructured substrates, bio-functionalization, and surface-enhanced Raman spectroscopy (SERS) characterization, are reported. SERS detection of surface-immobilized proteins, and probing of protein-ligand and aptamer-ligand binding is demonstrated. Biology Next-generation Sequencing of 16S Ribosomal RNA Gene Amplicons Sylvie Sanschagrin1, Etienne Yergeau1 1Energy, Mining and Environment, National Research Council Canada Characterizing microbial community has been a longstanding goal in environmental microbiology. Next-generation sequencing methods now allow for the characterization of microbial communities at an unprecedented depth with minimal cost and labor. We detail here our approach to sequence bacterial 16S ribosomal RNA genes using a benchtop sequencer. Neuroscience Culturing and Electrophysiology of Cells on NRCC Patch-clamp Chips Christophe Py1, Marzia Martina2, Robert Monette2, Tanya Comas2, Mike W. Denhoff1, Collin Luk3, Naweed I. Syed3, Geoff Mealing2 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. Immunology and Infection Multiplex Detection of Bacteria in Complex Clinical and Environmental Samples using Oligonucleotide-coupled Fluorescent Microspheres Tim J. Dumonceaux1, Jennifer R. Town1, Janet E. Hill2, Bonnie L. Chaban2, Sean M. Hemmingsen3 1Saskatoon Research Centre, Agriculture and Agri-Food Canada, 2Department of Veterinary Microbiology, University of Saskatchewan, 3Plant Biotechnology Institute, National Research Council of Canada We describe a multiplex method for the detection of microorganisms within a sample using oligonucleotide-coupled fluorescent beads. Amplicon from all organisms within a sample is hybridized to a panel of probe-coupled beads. A Luminex or Bio-Plex instrument is used to query each bead for bead type and hybridization signal.