University of Oslo

17 articles published in JoVE

• Chemistry

  
  Microsampling in Targeted Mass Spectrometry-Based Protein Analysis of Low-Abundance Proteins Huan Bao Ngo¹, Inger Oulie¹, Léon Reubsaet¹, Trine Grønhaug Halvorsen^{1 1}Department of Pharmacy, University of Oslo A protocol is presented for the determination of low-abundance biomarkers from dried serum samples exemplified with the biomarker progastrin-releasing peptide (ProGRP). Antibody-coated magnetic beads are used for the selective cleanup and enrichment of a proteotypic ProGRP peptide. The captured peptide is subsequently analyzed by liquid chromatography-tandem mass spectrometry.

• Medicine

  
  A Piglet Perinatal Asphyxia Model to Study Cardiac Injury and Hemodynamics after Cardiac Arrest, Resuscitation, and the Return of Spontaneous Circulation Eydis Oddsdottir Stenersen^1,2, Annette Olsen², Maria Melheim³, Rønnaug Solberg^3,4, Ingrid Dannevig⁵, Georg Schmölzer^6,7, Po-Yin Cheung^6,7, Britt Nakstad^8,9, Ola Didrik Saugstad^10,11, Arild Rønnestad^1,2, Anne Lee Solevåg^{2 1}Institute of Clinical Medicine, Faculty of Medicine, University of Oslo, ²Department of Neonatal Intensive Care, Division of Pediatric and Adolescent Medicine, Oslo University Hospital Rikshospitalet, ³Department of Pediatric Research, Division of Paediatric and Adolescent Medicine, Oslo University Hospital Rikshospitalet, ⁴Department of Pediatrics, Vestfold Hospital Trust, ⁵Department of Anesthesiology, Oslo University Hospital Rikshospitalet, ⁶Department of Pediatrics, Faculty of Medicine and Dentistry, University of Alberta, ⁷Centre for the Studies of Asphyxia and Resuscitation, Neonatal Research Unit, Royal Alexandra Hospital, ⁸Division of Paediatric and Adolescent Medicine, Institute of Clinical Medicine, University of Oslo, ⁹Department of Paediatrics and Adolescent Health, University of Botswana, ¹⁰Department of Pediatric Research, Oslo University Hospital, University of Oslo, ¹¹Department of Pediatrics, Robert H Lurie Medical Research Center, Northwestern University Chicago This piglet model involves surgical instrumentation, asphyxiation until the cardiac arrest, resuscitation, and post-resuscitation observation. The model allows for multiple sampling per animal, and by using continuous invasive arterial blood pressure, ECG, and non-invasive cardiac output monitoring, it provides knowledge about hemodynamics and cardiac pathophysiology in perinatal asphyxia and neonatal cardiopulmonary resuscitation.

• Bioengineering

  
  Subretinal Implantation of RPE on a Carrier in Minipigs: Guidelines for Preoperative Preparations, Surgical Techniques, and Postoperative Care Lyubomyr Lytvynchuk*^1,2, Zbynek Stranak*³, Hana Studenovska⁴, David Rais⁴, Štěpán Popelka⁴, Lucie Tichotová^5,6, Yaroslav Nemesh^5,6, Anastasiia Kolesnikova^5,6, Ruslan Nyshchuk^5,6, Anna Brymová^5,6, Zdeňka Ellederová⁵, Jana Čížková⁵, Jana Juhásová⁵, Štefan Juhás⁵, Pavla Jendelová⁷, Richárd Nagymihály⁸, Igor Kozak⁹, Slaven Erceg¹⁰, Susanne Binder¹¹, Brigitte Müller¹, Knut Stieger¹, Jan Motlik⁵, Goran Petrovski*⁸, Taras Ardan*^{5 1}Eye Clinic, Department of Ophthalmology, University Hospital Giessen and Marburg GmbH, ²Karl Landsteiner Institute for Retinal Research and Imaging, ³Department of Ophthalmology, University Hospital Kralovske Vinohrady and Third Faculty of Medicine, Charles University in Prague, ⁴Institute of Macromolecular Chemistry, Czech Academy of Sciences, ⁵Institute of Animal Physiology and Genetics, Czech Academy of Sciences, ⁶Department of Cell Biology, Faculty of Science, Charles University, ⁷Institute of Experimental Medicine, Czech Academy of Sciences, ⁸Center for Eye Research, Department of Ophthalmology, Oslo University Hospital and Institute of Clinical Medicine, Faculty of Medicine, University of Oslo, ⁹Moorfields Eye Hospitals UAE, ¹⁰Stem Cell Therapies in Neurodegenerative Diseases Lab, Research Center “Principe Felipe”, ¹¹Eye Center Donaustadt, Department of Ophthalmology, Sigmund Freud University The subretinal implantation of retinal pigmented epithelium (RPE) is one of the most promising approaches for the treatment of degenerative retinal diseases. However, the performance of preclinical studies on large-eye animal models remains challenging. This report presents guidelines for the subretinal transplantation of RPE on a cell carrier into minipigs.

• Chemistry

  
  Analyzing Melts and Fluids from Ab Initio Molecular Dynamics Simulations with the UMD Package Razvan Caracas^1,2, Anais Kobsch¹, Natalia V. Solomatova¹, Zhi Li¹, Francois Soubiran^1,3, Jean-Alexis Hernandez^{1,2 1}Ecole Normale Supérieure de Lyon, Laboratory of Geology of Lyon UMR5276, CNRS, ²Centre for Earth Evolution and Dynamics (CEED), University of Oslo, ³ Melts and fluids are ubiquitous vectors of mass transport in natural systems. We have developed an open-source package to analyze ab initio molecular-dynamics simulations of such systems. We compute structural (bonding, clusterization, chemical speciation), transport (diffusion, viscosity) and thermodynamic properties (vibrational spectrum).

• Behavior

  
  Using Facial Electromyography to Assess Facial Muscle Reactions to Experienced and Observed Affective Touch in Humans Anbjørn Ree¹, India Morrison², Håkan Olausson², Uta Sailer¹, Markus Heilig², Leah M. Mayo^{2 1}Department of Behavioral Sciences in Medicine, Institute of Basic Medical Sciences, Faculty of Medicine, University of Oslo, ²Center for Social and Affective Neuroscience, Department of Clinical and Experimental Medicine, Linköping University We describe a protocol to assess facial muscle activity in response to experienced and observed tactile stimulation using facial electromyography.

• Bioengineering

  
  Spontaneous Formation and Rearrangement of Artificial Lipid Nanotube Networks as a Bottom-Up Model for Endoplasmic Reticulum Elif Senem Köksal¹, Patrícia F. Belletati¹, Ganna Reint¹, Ragni Olsson¹, Kira D. Leitl¹, Ilayda Kantarci¹, Irep Gözen^{1,2,3 1}Centre for Molecular Medicine Norway, Faculty of Medicine, University of Oslo, ²Department of Chemistry, Faculty of Mathematics and Natural Sciences, University of Oslo, ³Department of Chemistry and Chemical Engineering, Chalmers University of Technology Solid-supported, protein-free, double phospholipid bilayer membranes (DLBM) can be transformed into complex and dynamic lipid nanotube networks and can serve as 2D bottom-up models of the endoplasmic reticulum.

• Cancer Research

  
  Phospho Flow Cytometry with Fluorescent Cell Barcoding for Single Cell Signaling Analysis and Biomarker Discovery Sigrid S. Skånland^{1,2 1}Centre for Molecular Medicine Norway (NCMM), Nordic EMBL Partnership, University of Oslo, ²K. G. Jebsen Centre for B cell malignancies and K. G. Jebsen Centre for Cancer Immunotherapy, University of Oslo Here, a protocol for medium- to high-throughput analysis of protein phosphorylation events at the cellular level is presented. Phospho flow cytometry is a powerful approach to characterize signaling aberrations, identify and validate biomarkers, and assess pharmacodynamics.

• Genetics

  
  Producing Gene Deletions in Escherichia coli by P1 Transduction with Excisable Antibiotic Resistance Cassettes Athanasios Saragliadis¹, Thomas Trunk¹, Jack C. Leo^{1 1}Evolution and Genetics, Department of Biosciences, University of Oslo Here we present a protocol for the use of pre-existing antibiotic resistance-cassette deletion constructs as a basis for making deletion mutants in other E. coli strains. Such deletion mutations can be mobilized and inserted into the corresponding locus of a recipient strain using P1 bacteriophage transduction.

• Neuroscience

  
  Preparation of a High-quality Primary Cell Culture from Fish Pituitaries Eirill Ager-Wick¹, Kjetil Hodne¹, Romain Fontaine¹, Kristine von Krogh¹, Trude M. Haug², Finn-Arne Weltzien^{1 1}Department of Basic Sciences and Aquatic Medicine, Faculty of Veterinary Medicine, Norwegian University of Life Sciences, ²Department of Oral Biology, Faculty of Dentistry, University of Oslo Here we describe a protocol to prepare and maintain primary pituitary cell cultures from medaka (Oryzias latipes). The optimized conditions in this protocol take important parameters such as temperature, osmolality, and pH into consideration by mimicking the physiological conditions of the fish, thereby enabling physiologically more meaningful results.

• Biology

  
  The Lactate Dehydrogenase Sequestration Assay — A Simple and Reliable Method to Determine Bulk Autophagic Sequestration Activity in Mammalian Cells Morten Luhr*¹, Paula Szalai*¹, Nikolai Engedal^{1 1}The Autophagy Team, Centre for Molecular Medicine Norway (NCMM), Nordic EMBL Partnership, University of Oslo Here a simple and well-validated protocol for measuring bulk autophagic sequestration activity in mammalian cells is described. The method is based on quantifying the proportion of lactate dehydrogenase (LDH) in sedimentable cell fractions compared to total cellular LDH levels.

• Neuroscience

  
  Visualization and Live Imaging of Oligodendrocyte Organelles in Organotypic Brain Slices Using Adeno-associated Virus and Confocal Microscopy Lauritz Hagen Kennedy¹, Johanne Egge Rinholm^{1,2 1}Division of Anatomy, Institute of Basic Medical Sciences, University of Oslo, ²Department of Microbiology, Oslo University Hospital Myelinating oligodendrocytes promote rapid action potential propagation and neuronal survival. Described here is a protocol for oligodendrocyte-specific expression of fluorescent proteins in organotypic brain slices with subsequent time-lapse imaging. Further, a simple procedure for visualizing unstained myelin is presented.

• Medicine

  
  The 4-vessel Sampling Approach to Integrative Studies of Human Placental Physiology In Vivo Ane M. Holme^1,2, Maia B. Holm^1,2, Marie C. P. Roland^1,3, Hildegunn Horne^1,2, Trond M. Michelsen^1,3, Guttorm Haugen^2,4, Tore Henriksen^{1,2 1}Department of Obstetrics, Oslo University Hospital, ²Institute of Clinical Medicine, University of Oslo, ³Norwegian Advisory Unit on Women's Health, Oslo University Hospital, ⁴Department of Fetal Medicine, Oslo University Hospital We present a detailed method to study human placental physiology in vivo at term. The method combines blood sampling from the incoming and outgoing vessels on the maternal and fetal sides of the placenta with ultrasound measurements of volume blood flow and placental tissue sampling.

• Immunology and Infection

  
  Flow Cytometry-based Assay for the Monitoring of NK Cell Functions Sara Tognarelli*^1,2, Benedikt Jacobs*^3,4, Nina Staiger^1,2, Evelyn Ullrich^{1,2 1}Childrens Hospital, Department of Pediatric Stem Cell Transplantation and Immunology, Johann Wolfgang Goethe-University, ²LOEWE Center for Cell and Gene Therapy, Johann Wolfgang Goethe-University, ³Institute for Cancer Research, Department of Cancer Immunology, Oslo University Hospital, Radiumhospital, ⁴The KG Jebsen Center for Cancer Immunotherapy, Institute of Clinical Medicine, University of Oslo A simple and reliable method is described here to analyze a set of NK cell functions such as degranulation, cytokine and chemokine production within different NK cell subsets.

• Medicine

  
  A Neonatal Mouse Spinal Cord Compression Injury Model Mark Züchner^1,2, Joel C. Glover^2,3, Jean-Luc Boulland^{2,3 1}Department of Neurosurgery, Oslo University Hospital, ²Norwegian Center for Stem Cell Research, Oslo University Hospital, ³Laboratory of Neural Development and Optical Recording (NDEVOR), Department of Physiology, Institute of Basic Medical Sciences, University of Oslo This article describes a method for generating a reproducible spinal cord compression injury (SCI) in the neonatal mouse. The model provides an advantageous platform for studying mechanisms of adaptive plasticity that underlie spontaneous functional recovery.

• Biology

  
  Cryo-electron Microscopy Specimen Preparation By Means Of a Focused Ion Beam Stefano Rubino^1,4, Petter Melin³, Paul Spellward², Klaus Leifer^{1 1}Department of Engineering Sciences, Uppsala University, ²Gatan Inc., ³Department of Microbiology, Swedish University of Agricultural Sciences, ⁴Physics Department, University of Oslo Cryo Electron Microscopes, either Scanning (SEM) or Transmission (TEM), are widely used for characterization of biological samples or other materials with a high water content¹. A SEM/Focused Ion Beam (FIB) is used to identify features of interest in samples and extract a thin, electron-transparent lamella for transfer to a cryo-TEM.

• Medicine

  
  Live Imaging of Drug Responses in the Tumor Microenvironment in Mouse Models of Breast Cancer Elizabeth S. Nakasone^1,2, Hanne A. Askautrud^2,3, Mikala Egeblad^{2 1}Watson School of Biological Sciences, ²Cold Spring Harbor Laboratory, ³Departments of Medical Genetics, University of Oslo and Oslo University Hospital We describe a method for imaging response to anti-cancer treatment in vivo and at single cell resolution.

• Biology

  
  Xenotransplantation of Human Stem Cells into the Chicken Embryo Jean-Luc Boulland¹, Gabor Halasi¹, Nedim Kasumacic¹, Joel C. Glover^{1,2 1}Department of Physiology, University of Oslo, ²Norwegian Center for Stem Cell Research, University of Oslo In this paper we present a method for transplanting human stem cells into various regions of the central nervous system of the chicken embryo. This provides an in vivo model for assessing the proliferation and differentiation of various types of human stem cells in embryonic tissue environments.