Technische Universitat Dresden View Institution's Website 23 articles published in JoVE Engineering Manufacturing Process for Non-Adhesive Super-Soft Vocal Fold Models Patrick Häsner1, Peter Birkholz1 1Institute of Acoustics and Speech Communication, Technische Universität Dresden This study demonstrates the manufacturing of non-sticky and super-soft vocal fold models by introducing a specific way to create the vocal fold layers, providing a detailed description of the manufacturing procedure, and characterizing the properties of the models. Immunology and Infection Observing Islet Function and Islet-Immune Cell Interactions in Live Pancreatic Tissue Slices Mollie K. Huber1, Denise M. Drotar2,3,4, Helmut Hiller1, Maria L. Beery1, Paul Joseph1, Irina Kusmartseva1, Stephan Speier2,3,4, Mark A. Atkinson1, Clayton E. Mathews1, Edward A. Phelps5 1Department of Pathology, Immunology, and Laboratory Medicine, University of Florida, 2Paul Langerhans Institute Dresden (PLID) of the Helmholtz Zentrum München at the University Clinic Carl Gustav Carus of Technische Universität Dresden, 3Institute of Physiology, Faculty of Medicine, Technische Universität Dresden, 4German Center for Diabetes Research (DZD), 5J. Crayton Pruitt Family Department of Biomedical Engineering, University of Florida This study presents the application of live pancreatic tissue slices to the study of islet physiology and islet-immune cell interactions. Engineering Cryogenic Liquid Jets for High Repetition Rate Discovery Science Chandra B. Curry*1,2, Christopher Schoenwaelder*1,3, Sebastian Goede4, Jongjin B. Kim1, Martin Rehwald5,6, Franziska Treffert1,7, Karl Zeil5, Siegfried H. Glenzer1, Maxence Gauthier1 1SLAC National Accelerator Laboratory, 2University of Alberta, 3Friedrich-Alexander-Universität Erlangen-Nürnberg, 4European XFEL, 5Helmholtz-Zentrum Dresden-Rossendorf, 6Technische Universität Dresden, 7Technische Universität Darmstadt This protocol presents the operation and principles of micron-scale cylindrical and planar cryogenic liquid jets. Until now, this system has been used as a high repetition rate target in laser-plasma experiments. Anticipated cross-disciplinary applications range from laboratory astrophysics to material science, and eventually next-generation particle accelerators. Genetics Universal and Efficient Electroporation Protocol for Genetic Engineering of Gastrointestinal Organoids Anne-Marlen Gaebler1, Alexander Hennig1, Katharina Buczolich1, Jürgen Weitz1, Thilo Welsch1, Daniel E. Stange1, Kristin Pape1 1Department of Visceral-, Thoracic and Vascular Surgery, University Hospital Carl Gustav Carus, Technische Universität Dresden This protocol describes an efficient electroporation method for the transfection of four different gastrointestinal organoid entities with larger plasmids (to the extent of 10 kB). It can be performed within one day and does not need extensive preparation or special, cost-intensive electroporation buffers. Medicine Ultrasonic-augmented Primary Adult Fibroblast Isolation Stephan R. Künzel1, Charlotte Schaeffer1, Karolina Sekeres1, Carola S. Mehnert1, Stephanie M. Schacht Wall1, Manja Newe1, Susanne Kämmerer1, Ali El-Armouche1 1Institute of Pharmacology and Toxicology, Faculty of Medicine Carl Gustav Carus, Technische Universität Dresden We present a protocol to isolate primary adult fibroblasts in an easy, fast and reliable way, performable by beginners (e.g., students). The procedure combines enzymatic tissue digestion and mechanical agitation with ultrasonic waves to obtain primary fibroblasts. The protocol can easily be adapted to specific experimental requirements (e.g., human tissue). Developmental Biology Adult Zebrafish Injury Models to Study the Effects of Prednisolone in Regenerating Bone Tissue Karina Geurtzen1, Franziska Knopf1,2 1CRTD – Center for Regenerative Therapies Dresden, TU Dresden, 2Center for Healthy Aging, TU Dresden Here, we describe 3 adult zebrafish injury models and their combined use with immunosuppressive drug treatment. We provide guidance on imaging of regenerating tissues and on detecting bone mineralization therein. Biology Analysis of Beta-cell Function Using Single-cell Resolution Calcium Imaging in Zebrafish Islets Sharan Janjuha1,2, Sumeet Pal Singh1, Nikolay Ninov1,2 1Center for Molecular and Cellular Bioengineering, TU Dresden, 2Paul Langerhans Institute Dresden of the Helmholtz Center Munich at the University Hospital Carl Gustav Carus of TU Dresden Beta-cell functionality is important for blood-glucose homeostasis, which is evaluated at single-cell resolution using a genetically encoded reporter for calcium influx. Neuroscience Modeling Amyloid-β42 Toxicity and Neurodegeneration in Adult Zebrafish Brain Prabesh Bhattarai1, Alvin Kuriakose Thomas2, Mehmet Ilyas Cosacak1, Christos Papadimitriou1, Violeta Mashkaryan3, Yixin Zhang2, Caghan Kizil1,3 1German Centre for Neurodegenerative Diseases (DZNE) Dresden within Helmholtz Association, 2B CUBE, Center for Molecular Bioengineering, Technische Universität Dresden, 3Center for Regenerative Therapies Dresden (CRTD), TU Dresden This protocol describes the synthesis, characterization, and injection of monomeric amyloid-β42 peptides for generating amyloid toxicity in adult zebrafish to establish an Alzheimer's disease model, followed by histological analyses and detection of aggregations. Cancer Research Isolation of Circulating Tumor Cells in an Orthotopic Mouse Model of Colorectal Cancer Susan Kochall1, May-Linn Thepkaysone1, Sebastián A. García1, Alexander M. Betzler1, Jürgen Weitz1,2,3, Christoph Reissfelder1, Sebastian Schölch1,2,3 1Department of Gastrointestinal, Thoracic and Vascular Surgery, Medizinische Fakultät Carl Gustav Carus, Technische Universität Dresden, 2German Cancer Consortium (DKTK), 3German Cancer Research Center (DKFZ) We describe the establishment of orthotopic colorectal tumors via injection of tumor cells or organoids into the cecum of mice and the subsequent isolation of circulating tumor cells (CTCs) from this model. Cancer Research A Genetically Engineered Mouse Model of Sporadic Colorectal Cancer Alexander M. Betzler1, Susan Kochall1, Linda Blickensdörfer2, Sebastian A. Garcia1, May-Linn Thepkaysone1, Lahiri K. Nanduri1, Michael H. Muders3, Jürgen Weitz1,4,5, Christoph Reissfelder1, Sebastian Schölch1,4,5 1Department of Gastrointestinal, Thoracic and Vascular Surgery, Medizinische Fakultät Carl Gustav Carus, Technische Universität Dresden, 2Department of General, Gastrointestinal and Transplant Surgery, University of Heidelberg, 3Department of Pathology, Medizinische Fakultät Carl Gustav Carus, Technische Universität Dresden, 4German Cancer Consortium (DKTK), 5German Cancer Research Center (DKFZ) A protocol for the establishment of a genetically engineered mouse model of colorectal cancer by segmental adeno-cre infection and its surveillance via high-resolution colonoscopy is presented. Bioengineering Transmission of Multiple Signals through an Optical Fiber Using Wavefront Shaping Daniel Haufe1, Nektarios Koukourakis1, Lars Büttner1, Jürgen W. Czarske1 1Faculty of Electrical and Computer Engineering, TU Dresden We demonstrate the transmission of multiple independent signals through a multimode fiber using wavefront shaping employing a single spatial light modulator. By modulating the wavefront for each signal individually, spatially separated foci are transmitted. Potential applications are multiplexed data transfer in communications engineering and endoscopic light delivery in biophotonics. Biology Isolation of CD146+ Resident Lung Mesenchymal Stromal Cells from Rat Lungs Jennifer J.P. Collins1,2, Marius A. Möbius1,3,4, Bernard Thébaud1,2,5 1Sinclair Centre for Regenerative Medicine, Ottawa Hospital Research Institute, 2University of Ottawa, 3Department of Neonatology and Pediatric Critical Care Medicine, Medical Faculty and University Hospital Carl Gustav Carus, Technische Universität Dresden, 4DFG Research Center and Cluster of Excellence for Regenerative Therapies (CRTD), Technische Universität, Dresden, 5 This protocol describes an isolation technique for obtaining primary lung resident mesenchymal stromal cells from rats, through the use of enzymatic digestion, density gradient separation, plastic adherence and CD146+ magnetic bead selection. Engineering Comprehensive Characterization of Extended Defects in Semiconductor Materials by a Scanning Electron Microscope Ellen Hieckmann1, Markus Nacke1, Matthias Allardt1, Yury Bodrov1, Paul Chekhonin2, Werner Skrotzki2, Jörg Weber1 1Institute of Applied Physics, Semiconductor Physics, Technische Universität Dresden, 2Institut für Strukturphysik, Technische Universität Dresden The optical, electrical, and structural properties of dislocations and of grain boundaries in semiconductor materials can be determined by experiments performed in a scanning electron microscope. Electron microscopy has been used to investigate cathodoluminescence, electron beam induced current, and diffraction of backscattered electrons. Engineering In Situ Time-dependent Dielectric Breakdown in the Transmission Electron Microscope: A Possibility to Understand the Failure Mechanism in Microelectronic Devices Zhongquan Liao1,2, Martin Gall1, Kong Boon Yeap1,3, Christoph Sander1, André Clausner1, Uwe Mühle1, Jürgen Gluch1, Yvonne Standke1, Oliver Aubel4, Armand Beyer4, Meike Hauschildt4, Ehrenfried Zschech1,2 1Fraunhofer Institute for Ceramic Technologies and Systems, 2Dresden Center for Nanoanalysis, Technische Universität Dresden, 3Globalfoundries Fab 8, 4Globalfoundries Fab 1 The time-dependent dielectric breakdown (TDDB) in on-chip interconnect stacks is one of the most critical failure mechanisms for microelectronic devices. This paper demonstrates the procedure of an in situ TDDB experiment in the transmission electron microscope, which opens a possibility to study the failure mechanism in microelectronic products. Immunology and Infection Isolation and Intravenous Injection of Murine Bone Marrow Derived Monocytes Martin Wagner1, Helen Koester1, Christian Deffge1, Soenke Weinert1, Johannes Lauf1, Alexander Francke2, Jerry Lee3, R. C. Braun- Dullaeus1, Joerg Herold1 1Department for Cardiology, Angiology and Pneumology, Otto von Guericke University Magdeburg, 2Herzzentrum Dresden, Universitätsklinikum an der Technischen Universität Dresden, Technische Universität Dresden, 3Department of Public Health and Primary Care, University of Cambridge Here we present a protocol that generates large amounts of murine monocytes from heterogeneous bone marrow for translational applications. In comparison to others, this new method helps reduce the number of sacrificed animals and lowers costs by avoiding expensive methods such as high gradient magnetic cell separation (MACS). Biology Reverse Genetic Morpholino Approach Using Cardiac Ventricular Injection to Transfect Multiple Difficult-to-target Tissues in the Zebrafish Larva Judith Konantz1, Christopher L. Antos1 1DFG-Center for Regenerative Therapies Dresden, Technische Universität Dresden An adaptable reverse genetic method for zebrafish to assess gene function during later stages of development and physiological homeostasis such as tissue regeneration using intraventricular injections of gene-specific morpholinos. Neuroscience One Mouse, Two Cultures: Isolation and Culture of Adult Neural Stem Cells from the Two Neurogenic Zones of Individual Mice Tara L. Walker1, Gerd Kempermann1,2 1Center for Regenerative Therapies Dresden, Technische Universität Dresden, 2German Center for Neurodegenerative Diseases (DZNE) Dresden Here we describe a detailed protocol for the simultaneous generation of neural precursor cell cultures, as either adherent monolayers or neurospheres, from the subventricular zone and dentate gyrus of individual adult mice. Medicine Subretinal Transplantation of MACS Purified Photoreceptor Precursor Cells into the Adult Mouse Retina Dominic Eberle*1, Tiago Santos-Ferreira*1, Sandra Grahl1, Marius Ader1 1CRTD / DFG-Research Center for Regenerative Therapies Dresden, Technische Universität Dresden Cell transplantation represents a strategy for the treatment of retinal degeneration characterized by photoreceptor loss. Here we describe a method for enrichment of transplantable photoreceptors and their subretinal grafting into adult mice. Neuroscience Growing Neural Stem Cells from Conventional and Nonconventional Regions of the Adult Rodent Brain Steven W. Poser1, Andreas Androutsellis-Theotokis1,2 1Department of Medicine, University of Dresden, 2Center for Regerative Therapies Dresden Neural stem cells harvested from the adult brain are increasing utilized in applications ranging from the basic research of nervous system development to exploring potential clinical applications in regenerative medicine. This makes rigorous control in the isolation and culturing conditions used to grow these cells critical to sound experimental outcomes. Neuroscience Micromanipulation of Gene Expression in the Adult Zebrafish Brain Using Cerebroventricular Microinjection of Morpholino Oligonucleotides Caghan Kizil1, Anne Iltzsche1, Jan Kaslin1, Michael Brand1 1DFG-Center for Regenerative Therapies Dresden, Cluster of Excellence (CRTD) and Biotechnology Center (BIOTEC) of the Technische Universität Dresden In this article, we demonstrate a method for manipulation of gene expression in the ventricular cells of the adult zebrafish telencephalon using antisense morpholino oligonucleotides. We present this method as an efficient and quick protocol that can be used for functional studies in the adult vertebrate brain. Medicine Improved Protocol For Laser Microdissection Of Human Pancreatic Islets From Surgical Specimens Dorothée Sturm1,2, Lorella Marselli3, Florian Ehehalt1,2, Daniela Richter1, Marius Distler2, Stephan Kersting1,2, Robert Grützmann2, Krister Bokvist4, Philippe Froguel5, Robin Liechti6, Anne Jörns7, Paolo Meda8, Gustavo Bruno Baretton9, Hans-Detlev Saeger2, Anke M. Schulte10, Piero Marchetti3, Michele Solimena1 1Molecular Diabetology, Paul Langerhans Institute Dresden, 2Department of GI-, Thoracic- and Vascular Surgery, University Hospital Carl Gustav Carus, University of Technology Dresden, 3Department of Endocrinology and Metabolism, Metabolic Unit University of Pisa, 4Labs DC0522, Lilly Corporate Center, 5Genomics, Faculty of Medicine Imperial College London, 6Vital-IT, SIB Swiss Institute of Bioinformatics, 7Clinical Biochemistry, Hannover Medical School, 8Cell Physiology and Metabolism, Medical School, University of Geneva, 9Department of Pathology, University Hospital Carl Gustav Carus, University of Technology Dresden, 10R&D DIAB Division / Translational Medicine, Sanofi-Aventis Laser microdissection is a technique that allows the recovery of selected cells from minute amounts of parenchyma. Here we describe a protocol for acquiring human pancreatic islets from surgical specimens to be used for transcriptomic studies. Our protocol improves the intrinsic autofluorescence of human beta cells, thus facilitating their collection. Medicine Isolation of Human Islets from Partially Pancreatectomized Patients Gregor Bötticher1, Dorothèe Sturm1, Florian Ehehalt1, Klaus P. Knoch2, Stephan Kersting1, Robert Grützmann1, Gustavo B. Baretton3, Michele Solimena2, Hans D. Saeger1 1Department of GI-, Thorax- and Vascular Surgery, University Hospital Carl Gustav Carus, University of Technology Dresden, 2Molecular Diabetology, Paul Langerhans Institute Dresden, 3Department of Pathology, University Hospital Carl Gustav Carus, University of Technology Dresden The supply of type 2 diabetic islets for research is insufficient. Here we share our protocol for isolating islets from patients undergoing partial pancreatectomy. This approach represents a unique venue for obtaining islets from type 2 diabetic and clinically matched non-diabetic subjects in adequate numbers for basic and clinical studies. Neuroscience Oral Administration of Rotenone using a Gavage and Image Analysis of Alpha-synuclein Inclusions in the Enteric Nervous System Francisco J. Pan-Montojo1, Richard H.W. Funk1 1Institute of Anatomy, Technische Universität Dresden Parkinson's disease has been related to the exposure to pesticides. Here we show a method to deliver pesticides using a gastric tube at the desired concentration and a method to analyze their effect in alpha-synuclein accumulation in the enteric nervous system.