University of Antwerp View Institution's Website 14 articles published in JoVE Biochemistry Design and Construction of an Experimental Setup to Enhance Mineral Weathering through the Activity of Soil Organisms Tullia Calogiuri1,2, Mathilde Hagens2, Jan Willem Van Groenigen1, Thomas Corbett3, Jens Hartmann4, Rick Hendriksen5, Iris Janssens6, Ivan A. Janssens7, Guillermo Ledesma Dominguez7, Grant Loescher4, Steven Mortier6, Anna Neubeck3, Harun Niron7, Reinaldy P. Poetra4, Lukas Rieder4, Eric Struyf7, Michiel Van Tendeloo8, Tom De Schepper6, Tim Verdonck9, Siegfried E. Vlaeminck8, Sara Vicca7, Alix Vidal1 1Soil Biology Group, Wageningen University & Research, 2Soil Chemistry and Chemical Soil Quality, Wageningen University & Research, 3Department of Earth Sciences, Uppsala University, 4Institute for Geology, Center for Earth System Research and Sustainability, University of Hamburg, 5Tupola, Wageningen University & Research, 6IDLab - Department of Computer Science, University of Antwerp - imec, 7Plants and Ecosystems (PLECO), Biology Department, University of Antwerp, 8Research Group of Sustainable Energy, Air and Water Technology, University of Antwerp, 9Department of Mathematics, University of Antwerp - imec Here we present the construction and operation of an experimental setup to enhance mineral weathering through the activity of soil organisms while concurrently manipulating abiotic variables known to stimulate weathering. Representative results from the functioning of the setup and sample analyses are discussed together with points for improvement. Cancer Research Multiparametric Tumor Organoid Drug Screening Using Widefield Live-Cell Imaging for Bulk and Single-Organoid Analysis Maxim Le Compte1, Edgar Cardenas De La Hoz2, Sofía Peeters1, Evelien Smits1, Filip Lardon1, Geert Roeyen1,3, Steve Vanlanduit2, Hans Prenen1,4, Marc Peeters1,4, Abraham Lin1,5, Christophe Deben1 1Center for Oncological Research (CORE), Integrated Personalized & Precision Oncology Network (IPPON), University of Antwerp, 2Industrial Vision Lab, University of Antwerp, 3Department of Hepatobiliary Transplantation and Endocrine Surgery, University Hospital Antwerp (UZA), 4Department of Oncology, University Hospital Antwerp (UZA), 5Plasma Lab for Applications in Sustainability and Medicine ANTwerp (PLASMANT), University of Antwerp This protocol describes a semi-automated method for medium- to high-throughput organoid drug screenings and microscope-agnostic, automated image analysis software to quantify and visualize multiparametric, single-organoid drug responses to capture intratumor heterogeneity. Biology An Experimental Approach to Investigating Effects of Artificial Light at Night on Free-Ranging Animals: Implementation, Results, and Directions for Future Research Melissa L. Grunst1, Andrea S. Grunst1, Rianne Pinxten1,2, Geert Eens1, Marcel Eens1 1Department of Biology, Behavioural Ecology and Ecophysiology Group, University of Antwerp, 2Faculty of Social Sciences, Antwerp School of Education, University of Antwerp Artificial light at night (ALAN) has wide-reaching biological effects. This article describes a system for manipulating ALAN inside nest boxes while monitoring behavior, consisting of LED lights coupled to a battery, timer, and audio-capable infrared video camera. Researchers could employ this system to explore many outstanding questions regarding the effects of ALAN on organisms. Medicine Sterile Pericarditis in Aachener Minipigs As a Model for Atrial Myopathy and Atrial Fibrillation Michiel R. L. Tubeeckx1, Steven Laga2, Christel Jacobs3, Marina Stroe4, Steven Van Cruchten4, Bo Goovaerts1, Jens Van fraeyenhove1, Hielko Miljoen5, Guido R. Y. De Meyer1, Gilles W. De Keulenaer1,6, Hein Heidbuchel5,7, Vincent F. M. Segers1,5 1Laboratory of Physiopharmacology, University of Antwerp, 2Department of Cardiac Surgery, University Hospital Antwerp, 3Department of Animal Welfare, University of Antwerp, 4Comparative Perinatal Development, University of Antwerp, 5Department of Cardiology, University Hospital Antwerp, 6Department of Cardiology, ZNA Middelheim Hospital, 7Research Group Cardiovascular Diseases, GENCOR, University of Antwerp We describe a sterile pericarditis model in minipigs to study atrial myopathy and atrial fibrillation (AF). We present surgical and anesthetic techniques, strategies for vascular access, and a protocol to study the inducibility of AF. Environment Protocol for Acute and Chronic Ecotoxicity Testing of the Turquoise Killifish Nothobranchius furzeri Charlotte Philippe1,2, Arnout F. Gregoir1, Eli S. J. Thoré1, Gudrun De Boeck2, Luc Brendonck1,3, Tom Pinceel1,4 1Animal Ecology, Global Change and Sustainable Development, University of Leuven, 2Systemic Physiological and Ecotoxicological Research, University of Antwerp, 3Water Research Group, Unit for Environmental Sciences and Management, North-West University, 4Centre for Environmental Management, University of the Free State In this work, we describe an acute, chronic and multigenerational bioassay to study the effects of single and combined stressors on the Turquoise killifish Nothobranchius furzeri. This protocol is designed to study life-history traits (mortality, growth, fecundity, weight) and critical thermal maximum. Medicine Non-invasive Assessments of Subjective and Objective Recovery Characteristics Following an Exhaustive Jump Protocol Erich Hohenauer1,2,3, Peter Clarys3, Jean-Pierre Baeyens2,3,4, Ron Clijsen1,2,3 1Department of Business Economics, Health and Social Care, University of Applied Sciences and Arts of Southern Switzerland, 2University College Physiotherapy "Thim van der Laan", 3Department of Movement and Sport Sciences, Vrije Universiteit Brussel, 4Faculty of Medicine and Health Sciences, University of Antwerp This protocol describes the procedure for non-invasive recovery assessments during a 72 h recovery period. This protocol induces muscular exhaustion of the frontal thighs using countermovement jumps and uses either local cold-cuff or thermoneutral-cuff application as a recovery modality. Biology Cellular Redox Profiling Using High-content Microscopy Tom Sieprath1,2, Tobias Corne1,2, Joke Robijns1, Werner J. H. Koopman3, Winnok H. De Vos1,2 1Laboratory of Cell Biology and Histology, Department of Veterinary Sciences, University of Antwerp, 2Cell Systems and Imaging Research Group (CSI), Department of Molecular Biotechnology, Ghent University, 3Department of Biochemistry , Radboud Institute for Molecular Life Sciences, Radboud University Medical Center This paper presents a high-content microscopy workflow for simultaneous quantification of intracellular ROS levels, as well as mitochondrial membrane potential and morphology – jointly referred to as mitochondrial morphofunction – in living adherent cells using the cell-permeant fluorescent reporter molecules 5-(and-6)-chloromethyl-2',7'-dichlorodihydrofluorescein diacetate, acetyl ester (CM-H2DCFDA) and tetramethylrhodamine methylester (TMRM). Biology Kinematic Analysis of Cell Division and Expansion: Quantifying the Cellular Basis of Growth and Sampling Developmental Zones in Zea mays Leaves Katrien Sprangers*1, Viktoriya Avramova*1, Gerrit T. S. Beemster1 1Department of Biology, University of Antwerp Quantifying cell division and expansion is of crucial importance to the understanding of whole-plant growth. Here, we present a protocol to calculate cellular parameters determining maize leaf growth rates and highlight the use of these data for investigating molecular growth regulatory mechanisms by directing developmental stage-specific sampling strategies. Neuroscience In Vitro Recording of Mesenteric Afferent Nerve Activity in Mouse Jejunal and Colonic Segments Sara Nullens1, Annemie Deiteren2, Wen Jiang3, Christopher Keating4, Hannah Ceuleers1, Sven Francque5, David Grundy3, Joris G. De Man1, Benedicte Y. De Winter1 1Laboratory of Experimental Medicine and Pediatrics, Division of Gastroenterology, University of Antwerp, 2Visceral Pain Group, Discipline of Medicine, University of Adelaide, 3Department of Biomedical Sciences, University of Sheffield, 4Department of Pharmacy, Pharmacology and Postgraduate Medicine, University of Hertfordshire, 5Department of Gastroenterology and Hepatology, Antwerp University Hospital Mesenteric afferent nerves convey information from the gastrointestinal tract towards the brain regarding normal homeostasis as well as pathophysiology. Gastrointestinal afferent nerve activity can be assessed by mounting isolated intestinal segments with attached afferent nerves into an organ bath, isolating the nerve, and assessing basal as well as stimulated activity. Medicine Exosomal miRNA Analysis in Non-small Cell Lung Cancer (NSCLC) Patients' Plasma Through qPCR: A Feasible Liquid Biopsy Tool Marco Giallombardo*1,2, Jorge Chacártegui Borrás*2,3, Marta Castiglia4, Nele Van Der Steen3, Inge Mertens5,6, Patrick Pauwels7,3, Marc Peeters8,3, Christian Rolfo2,3 1Department of Biopathology and Medical Biotechnology, Section of Biology and Genetics, University of Palermo, 2Phase I-Early Clinical Trials Unit, Oncology Department, Antwerp University Hospital (UZA), 3Center for Oncological Research (CORE), Antwerp University, 4Department of Surgical, Oncological and Oral Sciences, Section of Medical Oncology, University of Palermo, 5Flemish Institute for Technological Research (VITO), 6CORE, Campus Groenenborger, Antwerp University, 7Molecular Pathology, Pathology Department, Antwerp University Hospital (UZA), 8Oncology Department, Antwerp University Hospital (UZA) This protocol describes the feasibility to perform miRNA profiling in exosomes, released in plasma of NSCLC patients, through a commercial exosome isolation kit with Proteinase K and RNAse treatments, in order to avoid circulating miRNAs contamination and evaluate their biomarker features in NSCLC. Medicine Chronic Constriction Injury of the Rat's Infraorbital Nerve (IoN-CCI) to Study Trigeminal Neuropathic Pain Kristof Deseure1, Guy H. Hans2 1Translational Neurosciences, University of Antwerp, 2Multidisciplinary Pain Center, Antwerp University Hospital This manuscript describes a rodent model to study trigeminal neuropathic pain. The methods described include surgical procedures to perform a chronic constriction injury of the rat’s infraorbital nerve and the post-surgical behavioral tests to measure the changes in spontaneous and evoked behavior that are indicative of persistent pain and allodynia. Neuroscience Real-time Electrophysiology: Using Closed-loop Protocols to Probe Neuronal Dynamics and Beyond Daniele Linaro1, João Couto1, Michele Giugliano1 1Department of Biomedical Sciences, University of Antwerp Closed-loop protocols are becoming increasingly widespread in modern day electrophysiology. We present a simple, versatile and inexpensive way to perform complex electrophysiological protocols in cortical pyramidal neurons in vitro, using a desktop computer and a digital acquisition board. Behavior Functional Magnetic Resonance Imaging (fMRI) with Auditory Stimulation in Songbirds Lisbeth Van Ruijssevelt1, Geert De Groof1, Anne Van der Kant1, Colline Poirier1, Johan Van Audekerke1, Marleen Verhoye1, Annemie Van der Linden1 1Bio-Imaging Lab, University of Antwerp This article shows an optimized procedure for imaging of the neural substrates of auditory stimulation in the songbird brain using functional Magnetic Resonance Imaging (fMRI). It describes the preparation of the sound stimuli, the positioning of the subject and the acquisition and subsequent analysis of the fMRI data. Neuroscience Multimodal Imaging of Stem Cell Implantation in the Central Nervous System of Mice Nathalie De Vocht1,2, Kristien Reekmans1, Irene Bergwerf2, Jelle Praet1,2, Chloé Hoornaert1, Debbie Le Blon1, Jasmijn Daans1, Zwi Berneman1, Annemie Van der Linden2, Peter Ponsaerts1 1Laboratory of Experimental Hematology, University of Antwerp, 2Bio Imaging Lab, University of Antwerp This article describes an optimized sequence of events for multimodal imaging of cellular grafts in rodent brain using: (i) in vivo bioluminescence and magnetic resonance imaging, and (ii) post mortem histological analysis. Combining these imaging modalities on a single animal allows cellular graft evaluation with high resolution, sensitivity and specificity.