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Technische Universität Dresden

19 ARTICLES PUBLISHED IN JoVE

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Neuroscience

Oral Administration of Rotenone using a Gavage and Image Analysis of Alpha-synuclein Inclusions in the Enteric Nervous System
Francisco J. Pan-Montojo 1, Richard H.W. Funk 1
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.

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Medicine

Subretinal Transplantation of MACS Purified Photoreceptor Precursor Cells into the Adult Mouse Retina
Dominic Eberle *1, Tiago Santos-Ferreira *1, Sandra Grahl 1, Marius Ader 1
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.

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Neuroscience

One Mouse, Two Cultures: Isolation and Culture of Adult Neural Stem Cells from the Two Neurogenic Zones of Individual Mice
Tara L. Walker 1, Gerd Kempermann 1,2
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.

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Biology

Reverse Genetic Morpholino Approach Using Cardiac Ventricular Injection to Transfect Multiple Difficult-to-target Tissues in the Zebrafish Larva
Judith Konantz 1, Christopher L. Antos 1
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.

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Immunology and Infection

Isolation and Intravenous Injection of Murine Bone Marrow Derived Monocytes
Martin Wagner 1, Helen Koester 1, Christian Deffge 1, Soenke Weinert 1, Johannes Lauf 1, Alexander Francke 2, Jerry Lee 3, R. C. Braun- Dullaeus 1, Joerg Herold 1
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).

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Engineering

In Situ Time-dependent Dielectric Breakdown in the Transmission Electron Microscope: A Possibility to Understand the Failure Mechanism in Microelectronic Devices
Zhongquan Liao 1,2, Martin Gall 1, Kong Boon Yeap 1,3, Christoph Sander 1, André Clausner 1, Uwe Mühle 1, Jürgen Gluch 1, Yvonne Standke 1, Oliver Aubel 4, Armand Beyer 4, Meike Hauschildt 4, Ehrenfried Zschech 1,2
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.

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Biology

Isolation of CD146+ Resident Lung Mesenchymal Stromal Cells from Rat Lungs
Jennifer J.P. Collins 1,2, Marius A. Möbius 1,3,4, Bernard Thébaud 1,2,5
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, 5Children's Hospital of Eastern Ontario Research Institute

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.

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JoVE Core

Comprehensive Characterization of Extended Defects in Semiconductor Materials by a Scanning Electron Microscope
Ellen Hieckmann 1, Markus Nacke 1, Matthias Allardt 1, Yury Bodrov 1, Paul Chekhonin 2, Werner Skrotzki 2, Jörg Weber 1
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.

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Cancer Research

Isolation of Circulating Tumor Cells in an Orthotopic Mouse Model of Colorectal Cancer
Susan Kochall 1, May-Linn Thepkaysone 1, Sebastián A. García 1, Alexander M. Betzler 1, Jürgen Weitz 1,2,3, Christoph Reissfelder 1, Sebastian Schölch 1,2,3
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.

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Cancer Research

A Genetically Engineered Mouse Model of Sporadic Colorectal Cancer
Alexander M. Betzler 1, Susan Kochall 1, Linda Blickensdörfer 2, Sebastian A. Garcia 1, May-Linn Thepkaysone 1, Lahiri K. Nanduri 1, Michael H. Muders 3, Jürgen Weitz 1,4,5, Christoph Reissfelder 1, Sebastian Schölch 1,4,5
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.

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Neuroscience

Modeling Amyloid-β42 Toxicity and Neurodegeneration in Adult Zebrafish Brain
Prabesh Bhattarai 1, Alvin Kuriakose Thomas 2, Mehmet Ilyas Cosacak 1, Christos Papadimitriou 1, Violeta Mashkaryan 3, Yixin Zhang 2, Caghan Kizil 1,3
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.

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Medicine

Ultrasonic-augmented Primary Adult Fibroblast Isolation
Stephan R. Künzel 1, Charlotte Schaeffer 1, Karolina Sekeres 1, Carola S. Mehnert 1, Stephanie M. Schacht Wall 1, Manja Newe 1, Susanne Kämmerer 1, Ali El-Armouche 1
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).

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Genetics

Universal and Efficient Electroporation Protocol for Genetic Engineering of Gastrointestinal Organoids
Anne-Marlen Gaebler 1, Alexander Hennig 1, Katharina Buczolich 1, Jürgen Weitz 1, Thilo Welsch 1, Daniel E. Stange 1, Kristin Pape 1
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.

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JoVE Core

Cryogenic Liquid Jets for High Repetition Rate Discovery Science
Chandra B. Curry *1,2, Christopher Schoenwaelder *1,3, Sebastian Goede 4, Jongjin B. Kim 1, Martin Rehwald 5,6, Franziska Treffert 1,7, Karl Zeil 5, Siegfried H. Glenzer 1, Maxence Gauthier 1
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.

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Immunology and Infection

Observing Islet Function and Islet-Immune Cell Interactions in Live Pancreatic Tissue Slices
Mollie K. Huber 1, Denise M. Drotar 2,3,4, Helmut Hiller 1, Maria L. Beery 1, Paul Joseph 1, Irina Kusmartseva 1, Stephan Speier 2,3,4, Mark A. Atkinson 1, Clayton E. Mathews 1, Edward A. Phelps 5
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.

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Neuroscience

Isolation of Mouse Primary Microglia by Magnetic-Activated Cell Sorting in Animal Models of Demyelination
Hang Zhang 1, Sheng Yang 1, Man Chen 1, Dai-Shi Tian 1, Chuan Qin 1
1Department of Neurology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology

Here, we present a protocol to isolate and purify primary microglia in animal models of demyelinating diseases, utilizing columnar magnetic-activated cell sorting.

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Chemistry

Electrochemical Exfoliation to Produce High-Quality Black Phosphorus
Payam Hashemi 1,2, Davood Sabaghi 1, Sheng Yang 1, Ali Shaygan Nia 1,2, Xinliang Feng 1,2
1Center for Advancing Electronics Dresden (cfaed) and Department of Chemistry and Food Chemistry, Technische Universität Dresden, 2Department of Synthetic Materials and Functional Devices, Max Planck Institute of Microstructure Physics

We present a step-by-step procedure for electrochemical exfoliation of black phosphorus (BP), one of the most promising emerging 2D materials with applications in (opto)electronics, from its bulk crystals, as well as the morphological characterization by scanning electron microscopy, atomic force microscopy, and transmission electron microscopy.

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Engineering

Manufacturing Process for Non-Adhesive Super-Soft Vocal Fold Models
Patrick Häsner 1, Peter Birkholz 1
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.

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Neuroscience

Neuronavigated Focalized Transcranial Direct Current Stimulation Administered During Functional Magnetic Resonance Imaging
Filip Niemann *1, Alireza Shahbabaie *1, Sven Paßmann 1, Steffen Riemann 1, Robert Malinowski 1, Harund Kocataş 1, Leonardo M. Caisachana Guevara 1,2, Mohamed Abdelmotaleb 1, Daria Antonenko 1, Felix Blankenburg 3,4, Rico Fischer 2, Gesa Hartwigsen 5,6, Shu-Chen Li 7,8, Michael A. Nitsche 9,10,11, Axel Thielscher 12,13, Dagmar Timmann 14, Anna Fromm 1, Dayana Hayek 1, Ann-Kathrin Hubert 1, Andrew K. Martin 15,16, Alexander Hunold 17, Agnes Flöel *1,18, Marcus Meinzer *1
1Department of Neurology, University Medicine Greifswald, 2Department of Psychology, University Medicine Greifswald, 3Berlin School of Mind and Brain, Humboldt Universität zu Berlin, 4Neurocomputation and Neuroimaging Unit, Freie Universität Berlin, 5Wilhelm-Wundt-Institute for Psychology, Cognitive and Biological Psychology, Leipzig University, 6Lise Meitner Research Group Cognition and Plasticity, Max Planck Institute for Human Cognitive and Brain Sciences, 7Lifespan Developmental Neuroscience, Faculty of Psychology, Technische Universität Dresden, 8Centre for Tactile Internet with Human-in-the-Loop, Technische Universität Dresden, 9Department of Psychology and Neurosciences, Leibniz Research Centre for Working Environment and Human Factors, 10German Centre for Mental Health (DZPG), 11Clinic of Psychiatry and Psychotherapy, Protestant Hospital of Bethel Foundation, University Hospital OWL, Bielefeld University, 12Danish Research Centre for Magnetic Resonance, Centre for Functional and Diagnostic Imaging and Research, Copenhagen University Hospital Amager and Hvidovre, 13Department of Health Technology, Technical University of Denmark, 14Department of Neurology and Center for Translational Neuro- and Behavioral Sciences (C-TNBS), Essen University Hospital, 15Department of Psychology, University of Kent, 16Kent Medway Medical School, 17Institute of Biomedical Engineering and Informatics, Technische Universität Ilmenau, 18German Center for Neurodegenerative Diseases (DZNE Site Greifswald)

This protocol describes the method of neuronavigated electrode placement for focal, transcranial direct current stimulation (tDCS) administered during functional magnetic resonance imaging (fMRI).

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