University of Innsbruck View Institution's Website 8 articles published in JoVE Biochemistry Extraction and Purification of FAHD1 Protein from Swine Kidney and Mouse Liver Andreas Andric1, Eva Wagner1, Anne Heberle1, Max Holzknecht1, Alexander K. H. Weiss1 1Research Institute for Biomedical Aging Research, University of Innsbruck This protocol describes how to extract fumarylacetoacetate hydrolase domain-containing protein 1 (FAHD1) from swine kidney and mouse liver. The listed methods may be adapted to other proteins of interest and modified for other tissues. Biology Application of Membrane and Cell Wall Selective Fluorescent Dyes for Live-Cell Imaging of Filamentous Fungi Alexander Lichius1, Susanne Zeilinger1 1Department of Microbiology, University of Innsbruck Vital fluorescent dyes are essential tools for live-cell imaging analyses in modern fungal cell biology. This paper details the application of established and lesser-known fluorescent dyes for tracking plasma membrane dynamics, endo-/exocytosis and cell wall morphogenesis in filamentous fungi. Environment Laser-Induced Fluorescence Emission (L.I.F.E.) as Novel Non-Invasive Tool for In-Situ Measurements of Biomarkers in Cryospheric Habitats Klemens Weisleitner1,2, Lars Hunger3, Christoph Kohstall4, Albert Frisch5, Michael C. Storrie-Lombardi6, Birgit Sattler1,2 1Institute of Ecology, University of Innsbruck, 2Austrian Polar Research Institute, University of Vienna, 3BrainLinks-BrainTools, Bernstein Center Freiburg, 4Atom Science, Kasevich Lab, Stanford University, 5Institute of Experimental Physics, University of Innsbruck, 6Department of Physics, Extraterrestrial Vehicle Instruments Laboratory, Harvey Mudd College Carbon fluxes in the cryosphere are hardly assessed yet but are crucial regarding climate change. Here we show a novel prototype device that captures the phototrophic potential in supraglacial environments based on laser-induced fluorescence emission (L.I.F.E.) technology offering high spectral and spatial resolution data under in situ conditions. Biochemistry Medium Preparation for the Cultivation of Microorganisms under Strictly Anaerobic/Anoxic Conditions Andreas O. Wagner*1, Rudolf Markt*1, Mira Mutschlechner*1, Nina Lackner*1, Eva M. Prem*1, Nadine Praeg1, Paul Illmer1 1Department of Microbiology, Universität Innsbruck As obligate anaerobic organisms are unable to grow upon oxygen exposure, the use of anaerobic culturing techniques is indispensable. Here, we demonstrate a simple and effective method to cultivate a mixed culture derived from a biogas plant from media preparation to gas and volatile fatty acid quantification. Chemistry PTR-ToF-MS Coupled with an Automated Sampling System and Tailored Data Analysis for Food Studies: Bioprocess Monitoring, Screening and Nose-space Analysis Vittorio Capozzi1,2,3, Sine Yener1,2,4, Iuliia Khomenko1,5, Brian Farneti1, Luca Cappellin1, Flavia Gasperi1, Matteo Scampicchio2, Franco Biasioli1 1Department of Food Quality and Nutrition, Research and Innovation Centre, Fondazione Edmund Mach (FEM), 2Faculty of Science and Technology, Free University of Bolzano, 3Department of Agriculture, Food and Environmental Sciences, University of Foggia, 4Institute of Analytical Chemistry & Radiochemistry, Leopold-Franzens Universität Innsbruck, 5Institut für Ionenphysik und Angewandte Physik, Leopold-Franzens Universität Innsbruck Proton Transfer Reaction Time of Flight Mass Spectrometry allows high-sensitivity, rapid and non-invasive analysis of volatile organic compounds. To demonstrate its potential, we give three examples: lactic acid fermentation of yogurt (on-line bioprocess monitoring), different apple genotypes (large-scale screening), and retronasal space after drinking coffee (nosespace analysis). Developmental Biology Embryo Microinjection and Electroporation in the Chordate Ciona intestinalis Willi Kari1, Fan Zeng1, Lena Zitzelsberger1, Johannes Will1, Ute Rothbächer1 1Department of Evolution and Developmental Biology, Zoological Institute, University Innsbruck We present transient transgenesis and gene knockdown in Ciona intestinalis, a chordate sister group to vertebrates, using microinjection and electroporation techniques. Such methods facilitate functional genomics in this simple invertebrate that features rudimentary characteristics of vertebrates, including notochord and head sensory epithelia, and many orthologs of human disease associated genes. Environment The Use of High-resolution Infrared Thermography (HRIT) for the Study of Ice Nucleation and Ice Propagation in Plants Michael Wisniewski1, Gilbert Neuner2, Lawrence V. Gusta3 1U.S Department of Agriculture, Agricultural Research Service (USDA-ARS), Kearneysville, WV, 2Institute of Botany, University of Innsbruck, 3Department of Plant Science, University of Saskatechewan Here we present a protocol that allows one to visualize sites of ice formation and avenues of ice propagation in plants utilizing high resolution infrared thermography (HRIT). Biology Monitoring of Ubiquitin-proteasome Activity in Living Cells Using a Degron (dgn)-destabilized Green Fluorescent Protein (GFP)-based Reporter Protein Ruth Greussing1, Hermann Unterluggauer1, Rafal Koziel1, Andrea B. Maier2, Pidder Jansen-Dürr1 1Molecular and Cell Biology, Institute for Biomedical Aging Research, 2Department of Gerontology and Geriatrics, Netherlands Consortium for Healthy Aging, Leiden University Medical Center A method to monitor ubiquitin-proteasome activity in living cells is described. A degron-destabilized GFP- (GFP-dgn) and a stable GFP-dgnFS fusion protein are generated and transduced into the cell using a lentiviral expression vector. This technique allows to generate a stable GFP-dgn/GFP-dgnFS expressing cell line in which ubiquitin-proteasome activity can be easily assessed using epifluorescence or flow cytometry.