University of Zurich

43 articles published in JoVE

• Neuroscience

  
  Intracerebral Transplantation and In Vivo Bioluminescence Tracking of Human Neural Progenitor Cells in the Mouse Brain Rebecca Z. Weber^1,2, Chantal Bodenmann¹, Daniela Uhr¹, Kathrin J. Zürcher¹, Debora Wanner¹, Melanie Generali¹, Roger M. Nitsch¹, Ruslan Rust*^1,2, Christian Tackenberg*^{1,2 1}Institute for Regenerative Medicine, University of Zurich, ²Neuroscience Center Zurich, University of Zurich and ETH Zurich We describe the intraparenchymal transplantation of human neural progenitor cells transduced with a dual reporter vector expressing luciferase-green fluorescent protein (GFP) in the mouse brain. After transplantation, the luciferase signal is repeatedly measured using in vivo bioluminescence and GFP-expressing grafted cells identified in brain sections using fluorescence microscopy.

• Biochemistry

  
  Quaternary Structure Modeling Through Chemical Cross-Linking Mass Spectrometry: Extending TX-MS Jupyter Reports Hamed Khakzad^1,2, Swen Vermeul³, Lars Malmström^{4,5,6 1}Equipe Signalisation Calcique et Infections Microbiennes, Ecole Normale Supérieure Paris-Saclay, ²Institut National de la Santé et de la Recherche Médicale, ³Scientific IT Services, ETH Zurich, ⁴Institute for Computational Science, University of Zurich, ⁵S3IT, University of Zurich, ⁶Division of Infection Medicine, Department of Clinical Sciences Lund, Faculty of Medicine, Lund University Targeted cross-linking mass spectrometry creates quaternary protein structure models using mass spectrometry data acquired using up to three different acquisition protocols. When executed as a simplified workflow on the Cheetah-MS web server, the results are reported in a Jupyter Notebook. Here, we demonstrate the technical aspects of how the Jupyter Notebook can be extended for a more in-depth analysis.

• Medicine

  
  A Reproducible Intensive Care Unit-Oriented Endotoxin Model in Rats Jan Heil¹, Martin Schläpfer^{1,2 1}Institute of Physiology, University of Zurich, ²Institute of Anesthesiology, University Hospital of Zurich Here, we present a reproducible intensive care unit-oriented endotoxin model in rats.

• Developmental Biology

  
  Translating Ribosome Affinity Purification (TRAP) to Investigate Arabidopsis thaliana Root Development at a Cell Type-Specific Scale Martha Thellmann¹, Tonni Grube Andersen², Joop EM Vermeer^{1,3 1}Department of Plant and Microbial Biology, University of Zurich, ²Biophore - UNIL, ³Laboratory of Cell and Molecular Biology, Institute of Biology, University of Neuchâtel Translating ribosome affinity purification (TRAP) offers the possibility to dissect developmental programs with minimal processing of organs and tissues. The protocol yields high-quality RNA from cells targeted with a green fluorescent protein (GFP)-labeled ribosomal subunit. Downstream analysis tools, such as qRT-PCR or RNA-seq, reveal tissue and cell type-specific expression profiles.

• Immunology and Infection

  
  Infecting Mice with Malassezia spp. to Study the Fungus-Host Interaction Florian Sparber¹, Salomé LeibundGut-Landmann^{1 1}Section of Immunology, Vetsuisse Faculty, University of Zürich This protocol outlines the establishment of a mouse model for studying Malassezia-host interactions in the skin. It describes the cultivation of Malassezia in vitro, the infection of the murine skin with Malassezia, and the subsequent analysis of the inflammation and the fungal burden in the skin tissue.

• Neuroscience

  
  Delivery of Antibodies into the Murine Brain via Convection-enhanced Delivery Michal Beffinger¹, Linda Schellhammer¹, Stanislav Pantelyushin¹, Johannes vom Berg^{1 1}Institute of Laboratory Animal Science, University of Zurich Convection-enhanced delivery (CED) is a method enabling effective delivery of therapeutics into the brain by direct perfusion of large tissue volumes. The procedure requires the use of catheters and an optimized injection procedure. This protocol describes a methodology for CED of an antibody into a mouse brain.

• Biology

  
  Flow Cytometric Analysis of Extracellular Vesicles from Cell-conditioned Media Carolina Balbi*¹, Sara Bolis*¹, Giuseppe Vassalli^1,2,3, Lucio Barile^{1 1}Cellular and Molecular Cardiology Laboratory, Cardiocentro Ticino Foundation, Switzerland, ²Molecular Cardiology Institute, Dept. of Cardiology, University of Zürich, ³Faculty of Biomedical Science, Università Svizzera Italiana, Switzerland The protocol describes a reproducible method designed for use with cell culture supernatants to detect surface epitopes on small extracellular vesicles (EV). It utilizes specific EV immunoprecipitation using beads coupled with antibodies that recognize surface antigen CD9, CD63, and CD81. The method is optimized for downstream flow cytometry analysis.

• Behavior

  
  Virtual Reality Experiments with Physiological Measures Raphael P. Weibel¹, Jascha Grübel¹, Hantao Zhao¹, Tyler Thrash^1,2,3, Dario Meloni¹, Christoph Hölscher¹, Victor R. Schinazi^{1 1}Chair of Cognitive Science, ETH Zürich, ²Geographic Information Visualization and Analysis, University of Zürich, ³Digital Society Initiative, University of Zürich Virtual reality (VR) experiments can be difficult to implement and require meticulous planning. This protocol describes a method for the design and implementation of VR experiments that collect physiological data from human participants. The Experiments in Virtual Environments (EVE) framework is employed to accelerate this process.

• Behavior

  
  A Networked Desktop Virtual Reality Setup for Decision Science and Navigation Experiments with Multiple Participants Hantao Zhao¹, Tyler Thrash^1,2,3, Stefan Wehrli⁴, Christoph Hölscher¹, Mubbasir Kapadia⁵, Jascha Grübel¹, Raphael P. Weibel¹, Victor R. Schinazi^{1 1}Chair of Cognitive Science, ETH Zürich, ²Digital Society Initiative, University of Zürich, ³Department of Geography, University of Zürich, ⁴Decision Science Laboratory, ETH Zürich, ⁵Computer Science Department, Rutgers University This paper describes a method for conducting multi-user experiments on decision-making and navigation using a networked computer laboratory.

• Neuroscience

  
  Adeno-associated Virus-mediated Transgene Expression in Genetically Defined Neurons of the Spinal Cord Karen Haenraets*^1,2, Gioele W. Albisetti*¹, Edmund Foster¹, Hendrik Wildner^{1 1}Institute of Pharmacology and Toxicology, University of Zurich, ²Institute of Pharmaceutical Sciences, Swiss Federal Institute (ETH) Zurich Intraspinal injection of recombinase dependent recombinant adeno-associated virus (rAAV) can be used to manipulate any genetically labelled cell type in the spinal cord. Here we describe how to transduce neurons in the dorsal horn of the lumbar spinal cord. This technique enables functional interrogation of the manipulated neuron subtype.

• Developmental Biology

  
  Whole-mount Clearing and Staining of Arabidopsis Flower Organs and Siliques Afif Hedhly¹, Hannes Vogler¹, Christof Eichenberger¹, Ueli Grossniklaus^{1 1}Department of Plant and Microbial Biology, Zurich-Basel Plant Science Center, University of Zurich In this protocol, we describe techniques for the proper dissection of Arabidopsis flowers and siliques, some basic clearing techniques, and selected staining procedures for whole-mount observations of reproductive structures.

• Medicine

  
  Technique and Patient Selection Criteria of Right Anterior Mini-Thoracotomy for Minimal Access Aortic Valve Replacement Reza Tavakoli^1,2, Pascal Leprince¹, Max Gassmann², Peiman Jamshidi³, Nassrin Yamani⁴, Julien Amour⁵, Guillaume Lebreton^{1 1}Department of Cardiovascular and Thoracic Surgery, Pitié Salpêtrière University Hospital, Assistance Publique, Hôpitaux de Paris (APHP), Institut de Cardiologie, ²Institute of Veterinary Physiology and Zurich Center for Integrative Human Physiology, University of Zurich, ³Herzzentrum Hirslanden, Klinik St Anna, ⁴Department of Radiology, Canton Hospital Lucerne, ⁵Department of Anesthesiology and Intensive Care Unit, Pitié Salpêtrière University Hospital, Assistance Publique, Hôpitaux de Paris (APHP), Institut de Cardiologie The goal of this protocol is to describe in detail the technique of minimally invasive aortic valve replacement through a right anterior mini-thoracotomy and central aortic cannulation. This technique can potentially enhance patients' comfort and, by reducing post-operative morbidity, promote lowering the length of stay and global costs.

• Biology

  
  Cystometric and External Urethral Sphincter Measurements in Awake Rats with Implanted Catheter and Electrodes Allowing for Repeated Measurements Elena E. Foditsch^1,2, Karin Roider^1,2, Andrea M. Sartori^3,4,5, Thomas M. Kessler⁵, Sabik Raj Kayastha⁶, Ludwig Aigner², Marc P. Schneider^{7 1}Department of Urology, Spinal Cord Injury and Tissue Regeneration Center Salzburg, Paracelsus Medical University, ²Institute of Molecular Regenerative Medicine, Spinal Cord Injury and Tissue Regeneration Center Salzburg, Paracelsus Medical University, ³Brain Research Institute, University of Zürich, ⁴Department of Health Sciences and Technology, Swiss Federal Institute of Technology Zürich, ⁵Neuro-Urology, Spinal Cord Injury Center & Research, University of Zürich, Balgrist University Hospital, ⁶Department of Orthopaedics and Traumatology, Dhulikhel Hospital, Kathmandu University Hospital, ⁷Department of Urology, Inselspital, Bern University Hospital This protocol first describes the surgical procedure of the permanent implantation of a urinary bladder catheter combined with external urethral sphincter electrodes, and second, the measurement of the function of the urinary bladder and external urethral sphincter in implanted awake animals.

• Medicine

  
  Standardized Technique of Aortic Valve Re-implantation for Valve-sparing Aortic Root Replacement Reza Tavakoli^1,2, Guillaume Lebreton¹, Max Gassmann², Peiman Jamshidi³, Pascal Leprince^{1 1}Department of Cardiovascular and Thoracic Surgery, Pitie Salpetriere University Hospital, Assistance Publique, Hopitaux de Paris (APHP), Institut de Cardiologie, ²Institute of Veterinary Physiology and Zurich Center for Integrative Human Physiology, University of Zurich, ³Herzzentrum Hirslanden, Klinik St Anna Valve-sparing aortic root replacement has the advantage of preserving the patient's own aortic valve. The complexity of the reported techniques to date restricts their use to a limited number of cardiac surgeons. This protocol describes step-by-step a standardized technique reproducible by a greater number of cardiac surgeons.

• Developmental Biology

  
  Reprogramming Primary Amniotic Fluid and Membrane Cells to Pluripotency in Xeno-free Conditions Jaroslav Slamecka¹, Javier Laurini², Troy Shirley¹, Simon Philipp Hoerstrup³, Benedikt Weber^3,4,5, Laurie Owen¹, Steven McClellan^{1 1}Mitchell Cancer Institute, University of South Alabama, ²College of Medicine, University of South Alabama, ³Institute for Regenerative Medicine, University of Zurich, ⁴Department of Dermatology, University Hospital Zurich, ⁵Center for Applied Biotechnology and Molecular Medicine (CABMM), University of Zurich - Irchel Campus This protocol describes the reprogramming of primary amniotic fluid and membrane mesenchymal stem cells into induced pluripotent stem cells using a non-integrating episomal approach in fully chemically defined conditions. Procedures of extraction, culture, reprogramming, and characterization of the resulting induced pluripotent stem cells by stringent methods are detailed.

• Developmental Biology

  
  Correlative Super-resolution and Electron Microscopy to Resolve Protein Localization in Zebrafish Retina José M. Mateos¹, Gery Barmettler¹, Jana Doehner¹, Irene Ojeda Naharros², Bruno Guhl¹, Stephan C.F. Neuhauss², Andres Kaech¹, Ruxandra Bachmann-Gagescu^2,3, Urs Ziegler^{1 1}Center for Microscopy and Image Analysis, University of Zurich, ²Institute for Molecular Life Sciences, University of Zurich, ³Institute for Medical Genetics, University of Zurich This protocol describes the necessary steps to obtain subcellular protein localization results on zebrafish retina by correlating super-resolution light microscopy and scanning electron microscopy images.

• Medicine

  
  Technique of Minimally Invasive Transverse Aortic Constriction in Mice for Induction of Left Ventricular Hypertrophy Reza Tavakoli¹, Simona Nemska^1,2, Peiman Jamshidi³, Max Gassmann¹, Nelly Frossard^{2 1}Institute of Veterinary Physiology and Zurich Center for Integrative Human Physiology (ZIHP), University of Zurich, ²Laboratoire d'Innovation Thérapeutique, UMR7200, Université de Strasbourg - CNRS, ³Department of Cardiology, Canton Hospital Lucerne The aim of this protocol is to describe step-by-step the technique of minimally invasive transverse aortic constriction (TAC) in mice. By elimination of intubation and ventilation which are mandatory for the commonly used standard procedure, minimally invasive TAC simplifies the operative procedure and reduces the strain put on the animal.

• Medicine

  
  Rat Model of the Associating Liver Partition and Portal Vein Ligation for Staged Hepatectomy (ALPPS) Procedure Erik Schadde^1,2,3, Martin Hertl², Stefan Breitenstein³, Beatrice Beck-Schimmer^1,4, Martin Schläpfer^{1,4 1}Institute of Physiology - Center for Integrative Human Physiology, University of Zurich, ²Department of Surgery, Rush University Medical Center, ³Department of Surgery, Cantonal Hospital Winterthur, ⁴Institute of Anesthesiology, University and University Hospital Zurich Inducing rapid liver hypertrophy using Associating Liver Partition and Portal vein ligation for a Staged hepatectomy (ALPPS) has been proposed for resection of borderline resectable liver tumors. This model may elucidate mechanisms involved in rapid hypertrophy and allows testing of drugs that promote or block the acceleration of regeneration.

• Genetics

  
  Studying Cell Cycle-regulated Gene Expression by Two Complementary Cell Synchronization Protocols Aintzane Apraiz*¹, Jone Mitxelena*^2,3, Ana Zubiaga^{2 1}Department of Cell Biology and Histology, University of the Basque Country, UPV/EHU, ²Department of Genetics, Physical Anthropology and Animal Physiology, University of the Basque Country, UPV/EHU, ³Department of Molecular Mechanisms of Disease, University of Zurich We report two cell synchronization protocols that provide a context for studying events related to specific phases of the cell cycle. We show that this approach is useful for analyzing the regulation of specific genes in an unperturbed cell cycle or upon exposure to agents affecting the cell cycle.

• Medicine

  
  Full-root Aortic Valve Replacement by Stentless Aortic Xenografts in Patients with Small Aortic Roots Reza Tavakoli^1,2, Peiman Jamshidi³, Max Gassmann^{1,2 1}Institute of Veterinary Physiology, University of Zurich, ²Zurich Center of Integrative Human Physiology, University of Zurich, ³Department of Cardiology, Canton Hospital Lucerne Full-root aortic valve replacement by stentless aortic xenograft is a viable option in patients with small aortic roots. We describe, a technique for the full-root implantation of stentless aortic xenografts, with emphasis on the management of the proximal suture line and coronary anastomoses, and discuss its limitations and alternative options.

• Medicine

  
  Direct Re-implantation of Left Coronary Artery into the Aorta in Adults with Anomalous Origin of Left Coronary Artery from the Pulmonary Artery (ALCAPA) Reza Tavakoli¹, Peiman Jamshidi², Nassrin Yamani³, Max Gassmann^{1 1}Institute of Veterinary Physiology and Zurich Center of Integrative Human Physiology (ZIHP), University of Zurich, ²Department of Cardiology, Canton Hospital Lucerne, ³Department of Radiology, Canton Hospital Lucerne Surgical correction of ALCAPA is highly recommended, regardless of age or the degree of intercoronary collateralization. This protocol presents a technique for the direct re-implantation of adult-type ALCAPA into the aorta to re-establish the dual-coronary perfusion. Whenever feasible, direct re-implantation is preferred to other surgical correction techniques.

• Genetics

  
  Purification of High Molecular Weight Genomic DNA from Powdery Mildew for Long-Read Sequencing Joanna M. Feehan*^1,2, Katherine E. Scheibel*¹, Salim Bourras³, William Underwood⁴, Beat Keller³, Shauna C. Somerville^{1 1}Department of Plant and Microbial Biology, University of California Berkeley, ²John Innes Centre, Norwich Research Park, ³Department of Plant and Microbial Biology, University of Zürich, ⁴USDA-ARS Sunflower and Plant Biology Research Unit Described here is a method for the extraction, purification, and quality control of genomic DNA from the obligate biotrophic fungal pathogen, powdery mildew, for use in long-read genome sequencing.

• Biology

  
  The Mouse Isolated Perfused Kidney Technique Jan Czogalla¹, Frank Schweda², Johannes Loffing^{1 1}Institute of Anatomy, Swiss National Centre of Competence in Research Kidney, University of Zürich, ²Department of Physiology, University of Regensburg The mouse isolated perfused kidney (MIPK) is a technique for keeping a mouse kidney under ex vivo conditions perfused and functional for 1 hr. The buffers and surgical technique are described in detail.

• Biochemistry

  
  Characterization of Multi-subunit Protein Complexes of Human MxA Using Non-denaturing Polyacrylamide Gel-electrophoresis Patricia E. Nigg^1,2, Jovan Pavlovic^{1 1}Institute for Medical Virology, University of Zurich, ²Friedrich Miescher Institute for Biomedical Research This article describes a simple and rapid protocol to evaluate the oligomeric state of the dynamin-like GTPase MxA protein from lysates of human cells using a combination of non-denaturing PAGE with western blot analysis.

• Chemistry

  
  Characterization, Quantification and Compound-specific Isotopic Analysis of Pyrogenic Carbon Using Benzene Polycarboxylic Acids (BPCA) Daniel B. Wiedemeier¹, Susan Q. Lang², Merle Gierga³, Samuel Abiven¹, Stefano M. Bernasconi³, Gretchen L. Früh-Green³, Irka Hajdas⁴, Ulrich M. Hanke¹, Michael D. Hilf¹, Cameron P. McIntyre⁴, Maximilian P. W. Scheider¹, Rienk H. Smittenberg⁵, Lukas Wacker⁴, Guido L. B. Wiesenberg¹, Michael W. I. Schmidt^{1 1}Department of Geography, University of Zurich, ²Department of Earth and Ocean Sciences, University of South Carolina, ³Department of Earth Sciences, ETH Zurich, ⁴Laboratory of Ion Beam Physics, ETH Zurich, ⁵Department of Geological Sciences, Stockholm University We present the benzene polycarboxylic acid (BPCA) method for assessing pyrogenic carbon (PyC) in the environment. The compound-specific approach uniquely provides simultaneous information about the characteristics, quantity and isotopic composition (¹³C and ¹⁴C) of PyC.

• Immunology and Infection

  
  In Vitro Disassembly of Influenza A Virus Capsids by Gradient Centrifugation Sarah Stauffer^1,2, Firat Nebioglu³, Ari Helenius^{1 1}Institute of Biochemistry, ETH Zurich, ²Department of Biochemistry, University of Zurich, ³Institute for Molecular Health Sciences, ETH Zurich Disassembly of influenza A virus cores during virus entry into host cells is a multistep process. We describe an in vitro method to analyze the early stages of viral uncoating. In this approach, velocity gradient centrifugation is used to biochemically dissect the steps that initiate uncoating under defined conditions.

• Immunology and Infection

  
  Measuring Attachment and Internalization of Influenza A Virus in A549 Cells by Flow Cytometry Marie O. Pohl¹, Silke Stertz^{1 1}Institute of Medical Virology, University of Zurich We present a protocol describing a semi-quantitative method for measuring both, the attachment of influenza A virus to A549 cells, as well as the internalization of virus particles into the target cells by flow cytometry.

• Neuroscience

  
  Analysis of Developing Tooth Germ Innervation Using Microfluidic Co-culture Devices Pierfrancesco Pagella¹, Shayee Miran¹, Tim Mitsiadis^{1 1}Institute of Oral Biology, Unit of Orofacial Development and Regeneration, University of Zurich Co-cultures represent a valuable method to study the interactions between nerves and target tissues and organs. Microfluidic systems allow co-culturing ganglia and whole developing organs or tissues in different culture media, thus representing a valuable tool for the in vitro study of the crosstalk between neurons and their targets.

• Behavior

  
  Acquisition of a High-precision Skilled Forelimb Reaching Task in Rats Ajmal Zemmar*^1,2, Brigitte Kast*^1,2, Karin Lussi^1,2, Andreas R. Luft*³, Martin E. Schwab*^{1,2 1}Brain Research Institute, University of Zurich, ²Department of Biology and Department of Health Sciences and Technology, ETH Zurich, ³Clinical Neurorehabilitation, Department of Neurology, University of Zurich & University Hospital Zurich A paradigm is presented to analyze the acquisition of a high-precision skilled forelimb reaching task in rats.

• Developmental Biology

  
  An Enzymatic Method to Rescue Mesenchymal Stem Cells from Clotted Bone Marrow Samples Philipp Schlaefli*¹, Alessandro Bertolo*¹, Cherry Malonzo¹, Tobias Poetzel², Martin Baur³, Frank Steffen⁴, Jivko Stoyanov^{1 1}Swiss Paraplegic Research, ²Orthopaedics and Spinal Surgery, Swiss Paraplegic Centre, ³Department of Neurosurgery, Lucerne Cantonal Hospital (LUKS), ⁴Section of Small Animal Surgery/Neurology, Vetsuisse Faculty, University of Zurich Mesenchymal stem cells are usually obtained from bone marrow and require expansion culture. When samples clot before processing, a protocol using the (enzymatic) thrombolytic drug urokinase can be applied to degrade the clot. Thus, cells are released and available for expansion culture. This protocol provides a rapid and inexpensive alternative to resampling.

• Neuroscience

  
  Real-Time Impedance-based Cell Analyzer as a Tool to Delineate Molecular Pathways Involved in Neurotoxicity and Neuroprotection in a Neuronal Cell Line Zoya Marinova¹, Susanne Walitza¹, Edna Grünblatt^{1 1}University Clinics for Child and Adolescent Psychiatry, University of Zürich Improved in vitro neurotoxicity assays would aid the identification of new neuroprotective compounds. The utility of a real-time impedance-based cell analyzer to determine cytotoxicity and cytoprotection in neuronal cell lines and to delineate the involvement of second messenger pathways, thus gaining insight in the mechanism of neuroprotection is presented.

• Biology

  
  An Efficient Method for Quantitative, Single-cell Analysis of Chromatin Modification and Nuclear Architecture in Whole-mount Ovules in Arabidopsis Wenjing She¹, Daniel Grimanelli², Célia Baroux^{1 1}Institute of Plant Biology and Zürich-Basel Plant Science Center, University of Zürich, ²Institut de Recherche pour le Développement (UMR 232), Centre National de la Recherche Scientifique (ERL 5300), Université de Montpellier II We provide here an efficient and reliable protocol for immunostaining, Fluorescence in situ Hybridization, DNA staining followed by quantitative, high-resolution imaging in whole-mount Arabidopsis thaliana ovules. This method was successfully used to analyze chromatin modifications and nuclear architecture.

• Medicine

  
  Accuracy in Dental Medicine, A New Way to Measure Trueness and Precision Andreas Ender¹, Albert Mehl^{1 1}Division of Computer-assisted Restorative Dentistry, Center of Dental Medicine, University of Zürich Accuracy is a major demand in dental medicine. To verify accuracy, reference scanners are needed. This article presents a new reference scanner with an adjusted scanning method to acquire a broad variety of dental morphologies with high trueness and precision.

• Biology

  
  Mouse Genome Engineering Using Designer Nucleases Mario Hermann¹, Tomas Cermak², Daniel F. Voytas², Pawel Pelczar^{1 1}Institute of Laboratory Animal Science, University of Zurich, ²Department of Genetics, Cell Biology & Development and Center for Genome Engineering, University of Minnesota Designer nucleases such as zinc finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs) can be used to modify the genome of mouse preimplantation embryos by triggering both the nonhomologous end joining (NHEJ) and homologous recombination (HR) pathways. These advances enable the rapid generation of mice with precise genetic modifications.

• Chemistry

  
  Mizoroki-Heck Cross-coupling Reactions Catalyzed by Dichloro{bis[1,1',1''-(phosphinetriyl)tripiperidine]}palladium Under Mild Reaction Conditions Miriam Oberholzer¹, Christian M. Frech^{2 1}Institute of Inorganic Chemistry, University of Zürich, ²Institute of Chemistry & Biological Chemistry, Zürich University of Applied Sciences Dichloro{bis[1,1',1''-(phosphinetriyl)tripiperidine]}palladium [(P(NC5H10)3)2Pd(Cl)2] (1) is an easy accessible, cheap, and air stable, but highly active Heck catalyst with an excellent functional group tolerance that efficiently operates under mild reaction conditions to give the coupling products in very high yields.

• Biology

  
  Efficient and Rapid Isolation of Early-stage Embryos from Arabidopsis thaliana Seeds Michael T. Raissig¹, Valeria Gagliardini¹, Johan Jaenisch¹, Ueli Grossniklaus¹, Célia Baroux^{1 1}Institute of Plant Biology and Zürich-Basel Plant Science Center, University of Zürich We report an efficient and simple method to isolate embryos at early stages of development from Arabidopsis thaliana seeds. Up to 40 embryos can be isolated in 1 hr to 4 hr, depending on the downstream application. The procedure is suitable for transcriptome, DNA methylation, reporter gene expression, immunostaining and fluorescence in situ hybridization analyses.

• Behavior

  
  Perceptual and Category Processing of the Uncanny Valley Hypothesis' Dimension of Human Likeness: Some Methodological Issues Marcus Cheetham¹, Lutz Jancke^{1 1}Department of Neuropsychology, University of Zurich Investigation of the Uncanny Valley Hypothesis and affective experience requires an understanding of the hypothesis' dimension of human likeness (DHL). This protocol allows representation of the DHL and examination of categorical perception. Use of the same stimuli and fMRI to distinguish brain regions responsive to physical and category change is illustrated.

• Neuroscience

  
  In ovo Electroporation of miRNA-based Plasmids in the Developing Neural Tube and Assessment of Phenotypes by DiI Injection in Open-book Preparations Nicole H. Wilson¹, Esther T. Stoeckli^{1 1}Institute of Molecular Life Sciences, University of Zurich A method by which gene expression in the neural tube can be downregulated in a cell type-specific, traceable manner is described. We demonstrate how in ovo electroporation of microRNA-based plasmids that elicit spatiotemporally controlled RNA interference can be used to investigate commissural axon guidance in the developing neural tube.

• Medicine

  
  Roux-en-Y Gastric Bypass Operation in Rats Marco Bueter^1,2, Kathrin Abegg^2,3, Florian Seyfried⁴, Thomas A. Lutz^2,3, Carel W. le Roux^{4 1}Department of Surgery, University Hospital Zürich, ²Zürich Centre for Integrative Human Physiology, University of Zürich, ³Institute of Veterinary Physiology, Vetsuisse Faculty, University of Zürich, ⁴Imperial Weight Centre, Department of Investigative Medicine, Imperial College London Numerous studies using gastric bypass rat models have been recently conducted to uncover the underlying physiological mechanisms of Roux-en-Y gastric bypass operations. This article aims to demonstrate and discuss the technical and experimental details of our published gastric bypass rat model to understand advantages and limitations of this experimental tool.

• Medicine

  
  Implantation of Radiotelemetry Transmitters Yielding Data on ECG, Heart Rate, Core Body Temperature and Activity in Free-moving Laboratory Mice Nikola Cesarovic¹, Paulin Jirkof², Andreas Rettich², Margarete Arras^{1 1}Division of Surgical Research, University Hospital Zurich, ²Institute of Laboratory Animal Science, University of Zurich A surgical technique for implantation of commercially available telemetry transmitters used for continuous measurement of biopotential (one-lead ECG), heart rate, core body temperature and locomotor activity in freely moving mice is shown. Recommendations and protocols for post-operative care and pain relief, improving recovery, well being and survival rate are also presented.

• Neuroscience

  
  VisioTracker, an Innovative Automated Approach to Oculomotor Analysis Kaspar P. Mueller¹, Oliver D. R. Schnaedelbach², Holger D. Russig², Stephan C. F. Neuhauss^{1 1}Institute of Molecular Life Sciences, University of Zurich, ²TSE Systems GmbH The VisioTracker is an automated system for the quantitative analysis of visual performance of larval and small adult fish based on the recording of eye movements. It features full control over visual stimulus properties and real-time analysis, enabling high-throughput research in fields such as visual system development and function, pharmacology, neural circuit studies and sensorimotor integration.

• Neuroscience

  
  Combining Behavioral Endocrinology and Experimental Economics: Testosterone and Social Decision Making Christoph Eisenegger*¹, Michael Naef*^{2 1}Institute for Empirical Research in Economics, Laboratory for Social and Neural Systems Research, University of Zurich, ²Department of Economics, Royal Holloway, University of London The procedure described in this protocol shows that testosterone administration and folk beliefs about testosterone may be associated with directly opposed social behaviors.

• Biology

  
  Making MR Imaging Child's Play - Pediatric Neuroimaging Protocol, Guidelines and Procedure Nora M. Raschle^1,2, Michelle Lee¹, Roman Buechler¹, Joanna A. Christodoulou³, Maria Chang¹, Monica Vakil¹, Patrice L. Stering¹, Nadine Gaab^{1,3,4 1}Department of Developmental Medicine, Children’s Hospital Boston, ²Department of Neuropsychology, University of Zurich, ³Graduate School of Education, Harvard, ⁴Harvard Medical School Despite an increase in the use of structural and functional magnetic resonance imaging (fMRI) in humans, the study of young pediatric populations remains a challenge. We present a hands-on, step-by-step video protocol including guidelines for clinicians and researchers intending to perform (f)MRI in young children.