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Radboud University

11 ARTICLES PUBLISHED IN JoVE

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Neuroscience

Interfacing 3D Engineered Neuronal Cultures to Micro-Electrode Arrays: An Innovative In Vitro Experimental Model
Mariateresa Tedesco 1, Monica Frega 1,2, Sergio Martinoia 1, Mattia Pesce 3, Paolo Massobrio 1
1Department of Informatics, Bioengineering, Robotics and System Engineering (DIBRIS), University of Genova, 2Donders Institute for Brain, Cognition and Behaviour, Department of Cognitive Neuroscience, Radboud University Medical Center, 3Fondazione Istituto Italiano di Tecnologia (IIT)

In this work, a novel experimental model in which 3D neuronal cultures are coupled to planar Micro-Electrode Arrays (MEAs) is presented. 3D networks are built by seeding neurons in a scaffold made up of glass microbeads on which neurons grow and form interconnected 3D structures.

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Developmental Biology

Rapid Neuronal Differentiation of Induced Pluripotent Stem Cells for Measuring Network Activity on Micro-electrode Arrays
Monica Frega *1,2, Sebastianus H. C. van Gestel *3, Katrin Linda 2,3, Jori van der Raadt 3, Jason Keller 1,2, Jon-Ruben Van Rhijn 1,2, Dirk Schubert 1,2, Cornelis A. Albers 2,3,4, Nael Nadif Kasri 1,2,3
1Department of Cognitive Neurosciences, Radboudumc, 2Donders Institute for Brain, Cognition and Behaviour, Radboud University, 3Department of Human Genetics, Radboudumc, 4Department of Molecular Developmental Biology, Radboud University

We modify and implement a previously published protocol describing the rapid, reproducible, and efficient differentiation of human induced Pluripotent Stem Cells (hiPSCs) into excitatory cortical neurons12. Specifically, our modification allows for control of neuronal cell density and use on micro-electrode arrays to measure electrophysiological properties at the network level.

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Neuroscience

Two Algorithms for High-throughput and Multi-parametric Quantification of Drosophila Neuromuscular Junction Morphology
Anna Castells-Nobau *1, Bonnie Nijhof *1, Ilse Eidhof 1, Louis Wolf 2, Jolanda M. Scheffer-de Gooyert 1, Ignacio Monedero 3,4, Laura Torroja 3, Jeroen A.W.M. van der Laak 2,5, Annette Schenck *1
1Department of Human Genetics, Donders Institute for Brain, Cognition and Behaviour, Radboud University Medical Center, 2Microscopical Imaging Centre (MIC), Radboud University Medical Center, 3Department of Biology, Universidad Autónoma de Madrid, 4Department of Clinical and Experimental Medicine, Linköping University, 5Department of Pathology, Radboud University Medical Center

Two image analysis algorithms, "Drosophila NMJ Morphometrics" and "Drosophila NMJ Bouton Morphometrics" were created, to automatically quantify nine morphological features of the Drosophila neuromuscular junction (NMJ).

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Neuroscience

Drosophila Courtship Conditioning As a Measure of Learning and Memory
Tom S. Koemans 1,2,3, Cornelia Oppitz 4, Rogier A. T. Donders 5, Hans van Bokhoven 1,3, Annette Schenck 1,3, Krystyna Keleman 6, Jamie M. Kramer 7,8
1Department of Human Genetics, Radboud University Medical Center, 2Radboud Institute of Molecular Life Sciences, Radboud University, 3Donders Institute for Brain, Cognition, and Behaviour, Centre for Neuroscience, Radboud University, 4Research Institute of Molecular Pathology, Vienna, Austria, 5Department for Health Evidence, Radboud University Medical Center, 6Janelia Research Campus, Howard Hughes Medical Institute, 7Department of Physiology and Pharmacology, Schulich School of Medicine and Dentistry, Western University, 8Department of Biology, Faculty of Science, Western University

This protocol describes a Drosophila learning and memory assay called courtship conditioning. This classic assay is based on a reduction of male courtship behavior after sexual rejection by a non-receptive premated female. This natural form of behavioral plasticity can be used to test learning, short-term memory, and long-term memory.

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Neuroscience

High-throughput Analysis of Locomotor Behavior in the Drosophila Island Assay
Ilse Eidhof *1, Michaela Fenckova *1, Dei M. Elurbe 2, Bart van de Warrenburg 3, Anna Castells Nobau *1, Annette Schenck *1
1Department of Human Genetics, Donders Institute for Brain, Cognition and Behaviour, Radboud University Medical Center, 2Centre for Molecular and Biomolecular Informatics, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, 3Department of Neurology, Donders Institute for Brain, Cognition and Behaviour, Radboud University Medical Center

The island assay is a relatively new, cost-effective assay that can be used to evaluate the basic locomotor behavior of Drosophila melanogaster. This manuscript describes algorithms for automatic data processing and objective quantification of island assay data, making this assay a sensitive, high-throughput readout for large genetic or pharmacological screens.

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Engineering

In Situ Monitoring of the Accelerated Performance Degradation of Solar Cells and Modules: A Case Study for Cu(In,Ga)Se2 Solar Cells
Mirjam Theelen 1, Klaas Bakker 1, Henk Steijvers 1, Stefan Roest 2, Peter Hielkema 3, Nicolas Barreau 4, Erik Haverkamp 5,6
1TNO Solliance, Thin Film Technology, 2Eternal Sun, 3Hielkema Testequipment, 4Institut des Matériaux Jean Rouxel (IMN)-UMR 6502, Université de Nantes, CNRS, 5ReRa Solutions BV, 6Institute of Molecules and Materials, Radboud University

Two 'Combined Stress test with in situ measurement' setups, which allow real-time monitoring of accelerated degradation of solar cells and modules, were designed and constructed. These setups allow the simultaneous use of humidity, temperature, electrical biases, and illumination as independently controlled stress factors. The setups and various experiments executed are presented.

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Bioengineering

A Multilayer Microfluidic Platform for the Conduction of Prolonged Cell-Free Gene Expression
Ardjan J. van der Linden 1, Maaruthy Yelleswarapu 2, Pascal A. Pieters 1, Zoe Swank 3, Wilhelm T. S. Huck 2, Sebastian J. Maerkl 3, Tom F. A. de Greef 1,2
1Institute for Complex Molecular Systems, Department of Biomedical Engineering, Computational Biology Group, Eindhoven University of Technology, 2Institute for Molecules and Materials, Radboud University, 3Institute of Bioengineering, School of Engineering École Polytechnique Fédérale de Lausanne (EPFL)

The fabrication process of a PDMS-based, multilayer, microfluidic device that allows in vitro transcription and translation (IVTT) reactions to be performed over prolonged periods is described. Furthermore, a comprehensive overview of the hardware and software required to automate and maintain these reactions for prolonged durations is provided.

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Behavior

A Within-Subject Experimental Design using an Object Location Task in Rats
Gülberk Bayraktar *1,2, Kristoffer Højgaard *1,3, Luc Nijssen 1,4, Tomonori Takeuchi 1,5
1Danish Research Institute of Translational Neuroscience (DANDRITE), Department of Biomedicine, Aarhus University, 2Universitätsklinikum Würzburg, Institut für Klinische Neurobiologie, 3Translational Neuropsychiatry Unit, Department of Clinical Medicine, Aarhus University, 4Faculty of Science, Radboud University, 5Center for Proteins in Memory - PROMEMO, Danish National Research Foundation, Department of Biomedicine, Aarhus University

This protocol provides detailed steps for an object location task with four repetitions using the same cohort of rats. Weak and strong encoding can produce short- and long-term memories. The flexibility of the protocol with repetition can be beneficial for studies involving surgical operations by saving time and labor.

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Cancer Research

Multiplex Immunohistochemical Analysis of the Spatial Immune Cell Landscape of the Tumor Microenvironment
Mark A. J. Gorris 1,2, Evgenia Martynova *1,3, Mark W. D. Sweep *1,4, Iris A. E. van der Hoorn 1,5, Shabaz Sultan 1,3, Mike J. D. E. Claassens 1, Lieke L. van der Woude 1,2,6, Kiek Verrijp 1,2,6, Carl G. Figdor 1,3, Johannes Textor 1,3, I. Jolanda M. de Vries 1
1Department of Medical BioSciences, Radboudumc, 2Division of Immunotherapy, Oncode Institute, Radboudumc, 3Data Science, Institute for Computing and Information Sciences, Radboud University, 4Department of Medical Oncology, Radboudumc, 5Department of Pulmonary Diseases, Radboudumc, 6Department of Pathology, Radboudumc

This protocol describes in detail how immune cell characterization of the tumor microenvironment using multiplex immunohistochemistry is carried out.

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Neuroscience

The TD Drive: A Parametric, Open-Source Implant for Multi-Area Electrophysiological Recordings in Behaving and Sleeping Rats
Tim Schröder 1,2, Jacqueline van der Meij 1, Paul van Heumen 1,2, Anumita Samanta 1, Lisa Genzel 1
1Donders Institute for Brain, Cognition and Behaviour, Radboud University, 23D Neuro B.V.

Here, we present a unique, 3D-printable implant for rats, named TD Drive, capable of symmetric, bilateral wire electrode recordings, currently in up to ten distributed brain areas simultaneously.

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Neuroscience

The DREAM Implant: A Lightweight, Modular, and Cost-Effective Implant System for Chronic Electrophysiology in Head-Fixed and Freely Behaving Mice
Tim Schröder *1,2, Robert Taylor *3, Muad Abd El Hay 3, Abdellatif Nemri 2, Arthur França 1, Francesco Battaglia 1, Paul Tiesinga 1, Marieke L. Schölvinck *3, Martha N. Havenith *1,2,3
1Donders Institute for Brain, Cognition and Behaviour, Radboud University, 23D Neuro B.V., 3Zero-Noise Lab, Ernst-Strüngmann Institute for Neuroscience

Here, we introduce a lightweight, cost-effective probe implant system for chronic electrophysiology in rodents optimized for ease of use, probe recovery, experimental versatility, and compatibility with behavior.

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