University of Bergen 7 articles published in JoVE Bioengineering Real-Time Intravital Multiphoton Microscopy to Visualize Focused Ultrasound and Microbubble Treatments to Increase Blood-Brain Barrier Permeability Charissa Poon1,2, Melina Mühlenpfordt*3, Marieke Olsman*3, Spiros Kotopoulis4,5, Catharina de Lange Davies3, Kullervo Hynynen1,2,6 1Physical Sciences Platform, Sunnybrook Research Institute, 2Institute of Biomedical Engineering, University of Toronto, 3Department of Physics, Norwegian University of Science and Technology, 4Department of Clinical Medicine, University of Bergen, 5Exact Therapeutics AS, 6Department of Medical Biophysics, University of Toronto This protocol describes the surgical and technical procedures that enable real-time in vivo multiphoton fluorescence imaging of the rodent brain during focused ultrasound and microbubble treatments to increase blood-brain barrier permeability. Neuroscience Flow Cytometric Analysis of Multiple Mitochondrial Parameters in Human Induced Pluripotent Stem Cells and Their Neural and Glial Derivatives Kristina Xiao Liang*1,2, Anbin Chen*1,2,3,4, Cecilie Katrin Kristiansen1,2, Laurence A. Bindoff1,2 1Department of Clinical Medicine (K1), University of Bergen, 2Neuro-SysMed, Center of Excellence for Clinical Research in Neurological Diseases, Haukeland University Hospital, 3Department of Neurosurgery, Qilu Hospital and Institute of Brain and Brain-Inspired Science, Cheeloo College of Medicine, Shandong University, 4Shandong Key Laboratory of Brain Function Remodeling This study reports a novel approach to measure multiple mitochondrial functional parameters based on flow cytometry and double staining with two fluorescent reporters or antibodies to detect changes in mitochondrial volume, mitochondrial membrane potential, reactive oxygen species level, mitochondrial respiratory chain composition, and mitochondrial DNA. Biology Assessing Mineral Availability in Fish Feeds using Complementary Methods Demonstrated with the Example of Zinc in Atlantic Salmon Marta S. Silva1,2, Thea Stewart4, Heidi Amlund1,3, Jens J. Sloth1,3, Pedro Araujo1, Erik-Jan Lock1, Christer Hogstrand4, Robin Ørnsrud1, Rune Waagbø1,2, Antony Jesu Prabhu1 1Institute of Marine Research, 2Department of Biological Sciences, University of Bergen, 3National Food Institute, Technical University of Denmark, 4 This article explains in detail a systematic approach to assess micro-mineral availability in Atlantic salmon. The methodology includes tools and models with increasing biological complexity: (1) chemical speciation analysis, (2) in vitro solubility, (3) uptake studies in cell lines, and (4) in vivo fish studies. Cancer Research A 3D Spheroid Model for Glioblastoma Joris Guyon1,2, Laetitia Andrique1,2, Nadège Pujol1,2, Gro Vatne Røsland3,4, Gaelle Recher1,5,6, Andreas Bikfalvi1,2, Thomas Daubon1,2,4 1University Bordeaux, LAMC, 2INSERM U1029, University Bordeaux, 3Department of Biomedicine, University of Bergen, 4CNRS, IBGC UMR5095, 5LP2N, CNRS UMR 5298, IOA, 6Institut d'Optique Graduate School, IOA Here, we describe an easy-to-use invasion assay for glioblastoma. This assay is suitable for glioblastoma stem-like cells. A Fiji macro for easy quantification of invasion, migration, and proliferation is also described. Genetics Transcription Start Site Mapping Using Super-low Input Carrier-CAGE Nevena Cvetesic1,2, Elena Pahita1,2, Boris Lenhard1,2,3 1Institute of Clinical Sciences, Faculty of Medicine, Imperial College London, 2MRC London Institute of Medical Sciences, 3Sars International Centre for Marine Molecular Biology, University of Bergen Cap Analysis of Gene Expression (CAGE) is a method for genome-wide quantitative mapping of mRNA 5’ends to capture RNA polymerase II transcription start sites at a single-nucleotide resolution. This work describes a low-input (SLIC-CAGE) protocol for generation of high-quality libraries using nanogram-amounts of total RNA. Genetics Multi-locus Variable-number Tandem-repeat Analysis of the Fish-pathogenic Bacterium Yersinia ruckeri by Multiplex PCR and Capillary Electrophoresis Snorre Gulla1, Saima Nasrin Mohammad1, Duncan John Colquhoun1,2 1Fish Health Research Group, Norwegian Veterinary Institute, 2University of Bergen The Multi-Locus Variable-number tandem-repeat Analysis (MLVA) assay presented here enables inexpensive, robust and portable high-resolution genotyping of the fish-pathogenic bacterium Yersinia ruckeri. Starting from pure cultures, the assay employs multiplex PCR and capillary electrophoresis to produce ten-loci MLVA profiles for downstream applications. Biochemistry Investigating Protein Sequence-structure-dynamics Relationships with Bio3D-web Shashank Jariwala*1, Lars Skjærven*2, Xin-Qiu Yao1, Barry J. Grant1 1Department of Computational Medicine and Bioinformatics, University of Michigan Medical School, 2Department of Biomedicine, University of Bergen A protocol for the online investigation of protein sequence-structure-dynamics relationships using Bio3D-web is presented.
