KTH Royal Institute of Technology View Institution's Website 8 articles published in JoVE Developmental Biology Generation of a Human iPSC-Based Blood-Brain Barrier Chip Srikanth Jagadeesan1,2,3, Michael J. Workman4, Anna Herland5,6, Clive N. Svendsen4, Gad D. Vatine1,2,3 1The Department of Physiology and Cell Biology, Faculty of Health Sciences, Ben-Gurion University of the Negev, 2The Regenerative Medicine and Stem Cell (RMSC) Research Center, Ben-Gurion University of the Negev, 3The Zlotowski Center for Neuroscience, Ben-Gurion University of the Negev, 4The Board of Governors Regenerative Medicine Institute, Cedars-Sinai Medical Center, 5Division of Micro and Nanosystems, KTH Royal Institute of Technology, 6AIMES, Department of Neuroscience, Karolinska Institutet The blood-brain barrier (BBB) is a multicellular neurovascular unit tightly regulating brain homeostasis. By combining human iPSCs and organ-on-chip technologies, we have generated a personalized BBB chip, suitable for disease modeling and CNS drug penetrability predictions. A detailed protocol is described for the generation and operation of the BBB chip. Chemistry Rapid Nanoprobe Signal Enhancement by In Situ Gold Nanoparticle Synthesis Jorge T. Dias1, Gustav Svedberg1, Mats Nystrand2, Helene Andersson-Svahn1, Jesper Gantelius1 1Division of Proteomics and Nanobiotechnology, Science for Life Laboratory, KTH Royal Institute of Technology, 2Global Research and Development, Thermo Fisher Scientific IDD In this work, a protocol for signal enhancement of nanoprobe-based biosensing is presented. The protocol is based on the reduction of chloroauric acid onto the surface of existing nanoprobes that consist of gold, silver, silica or iron-oxide nanoparticles. Biochemistry Method to Visualize and Analyze Membrane Interacting Proteins by Transmission Electron Microscopy Ramakrishnan B. Kumar1, Lin Zhu2, Hans Hebert1,2, Caroline Jegerschöld1,2 1Department of Biosciences and Nutrition, Karolinska Institutet, 2School of Technology and Health, KTH Royal Institute of Technology Many proteins perform their function when attached to membrane surfaces. The binding of extrinsic proteins on nanodisc membranes can be indirectly imaged by transmission electron microscopy. We show that the characteristic stacking (rouleau) of nanodiscs induced by the negative stain sodium phosphotungstate is prevented by the binding of extrinsic protein. Biology Scanning Electron Microscopy (SEM) Protocols for Problematic Plant, Oomycete, and Fungal Samples M. Angélica Bello1, Yolanda Ruiz-León2, J. Vladimir Sandoval-Sierra3, Svetlana Rezinciuc4, Javier Diéguez-Uribeondo3 1Biodiversity and Conservation Department, Real Jardín Botánico, CSIC, 2Research Support Unit, Real Jardín Botánico, CSIC, 3Mycology Department, Real Jardín Botánico, CSIC, 4Division of Glycoscience, AlbaNova University Center, Royal Institute of Technology (KTH) Problems in the processing of biological samples for scanning electron microscopy observation include cell collapse, treatment of samples from wet microenvironments and cell destruction. Low-cost and relatively rapid protocols suited for preparing challenging samples such as floral meristems, oomycete cysts, and fungi (Agaricales) are compiled and detailed here. Bioengineering Film Extrusion of Crambe abyssinica/Wheat Gluten Blends Mikael Gällstedt1, Henrik Pettersson2, Therese Johansson2, William R. Newson3, Eva Johansson3, Mikael S. Hedenqvist4 1SIG Combibloc, 2Innventia, 3Department of Plant Breeding, The Swedish University of Agricultural Sciences, 4Fibre and Polymer Technology, KTH Royal Institute of Technology The side stream from plant oil production of Crambe abyssinica has limited value. The purpose of this study was to find methods for extruding materials based on this side stream, demonstrating that products with a higher value can be produced. The extrudates were found to have promising properties. Chemistry Unraveling Entropic Rate Acceleration Induced by Solvent Dynamics in Membrane Enzymes Charlotte Kürten1, Per-Olof Syrén1 1Science for Life Laboratory, School of Biotechnology, KTH Royal Institute of Technology Channels for the transportation of water molecules in enzymes influence active site solvation and catalysis. Herein we present a protocol for the engineering of these additional catalytic motifs based on in silico computer modeling and experiments. This will enhance our understanding of the influence of solvent dynamics on enzyme catalysis. Environment Transcript and Metabolite Profiling for the Evaluation of Tobacco Tree and Poplar as Feedstock for the Bio-based Industry Colin Ruprecht1, Takayuki Tohge1, Alisdair Fernie1, Cara L. Mortimer2, Amanda Kozlo2, Paul D. Fraser2, Norma Funke1, Igor Cesarino3,4, Ruben Vanholme3,4, Wout Boerjan3,4, Kris Morreel3,4, Ingo Burgert5,6, Notburga Gierlinger5,6, Vincent Bulone7, Vera Schneider8, Andrea Stockero8, Juan Navarro-Aviñó9, Frank Pudel10, Bart Tambuyser11, James Hygate12, Jon Bumstead13, Louis Notley13, Staffan Persson1,14 1Max Planck Institute for Molecular Plant Physiology, 2School of Biological Sciences, Plant Molecular Science, Centre for Systems and Synthetic Biology, Royal Holloway, University of London, 3Department of Plant Systems Biology, VIB, 4Department of Plant Biotechnology and Bioinformatics, UGhent, 5Institute for Building Materials, ETH Zurich, 6Applied Wood Materials, EMPA, 7Division of Glycoscience, School of Biotechnology, AlbaNova University Center, Royal Institute of Technology (KTH), 8European Research and Project Office GmbH, 9ABBA Gaia S.L., 10Pflanzenöltechnologie, 11Capax Environmental Services, 12Green Fuels, 13Neutral Consulting Ltd, 14Plant Cell Biology Research Centre, School of Botany, University of Melbourne Plant biomass offers a renewable resource for multiple products, including fuel, feed, food, and a variety of materials. In this paper we investigate the properties of tobacco tree (Nicotiana glauca) and poplar as suitable sources for a biorefinery pipeline. Biology Orthogonal Protein Purification Facilitated by a Small Bispecific Affinity Tag Johan Nilvebrant1, Tove Alm1, Sophia Hober1 1School of Biotechnology, Department of Proteomics, Royal Institute of Technology A novel and highly efficient two-step affinity chromatography protocol has been developed and is described in detail. The method is based on a small purification tag with two inherent affinities and is applicable to a wide range of target proteins with different properties.