Academic Medical Center Amsterdam View Institution's Website 10 articles published in JoVE Cancer Research Laparoscopic Pancreatoduodenectomy With Modified Blumgart Pancreaticojejunostomy Matteo De Pastena1,2, Jony van Hilst1, Thijs de Rooij1, Olivier R Busch1, Michael F Gerhards3, Sebastiaan Festen3, Marc G Besselink1 1Department of Surgery, Cancer Center Amsterdam, Academic Medical Center, 2General and Pancreatic Surgery Department, Pancreas Institute, University and Hospital Trust of Verona, 3Department of Surgery, OLVG Laparoscopic pancreatoduodenctomy (LPD) may offer advantages over open pancreatoduodenectomy, including early postoperative mobilization, less delayed gastric emptying and a shorter hospital stay. However, LPD is technically challenging and not well-standardized, especially regarding the pancreatic anastomosis. We describe a standardized technique for the pancreatic anastomosis during LPD: modified Blumgart pancreaticojejunostomy. Biochemistry Detection and Visualization of DNA Damage-induced Protein Complexes in Suspension Cell Cultures Using the Proximity Ligation Assay Mahnoush Bahjat1, Timon A. Bloedjes1, Amélie van der Veen1, Guus de Wilde1, Chiel Maas1, Jeroen E. J. Guikema1 1Department of Pathology, Academic Medical Center, University of Amsterdam, Lymphoma and Myeloma Center Amsterdam (LYMMCARE) Here, it is demonstrated how the in situ Proximity Ligation Assay (PLA) can be used to detect and visualize the direct protein-protein interactions between ATM and p53 in suspension cell cultures exposed to genotoxic stress. Developmental Biology A Submerged Filter Paper Sandwich for Long-term Ex Ovo Time-lapse Imaging of Early Chick Embryos Manuel Schmitz*1, Ben K. A. Nelemans*1, Theodoor H. Smit2 1Department of Orthopaedic Surgery, VU University Medical Center Amsterdam, MOVE Research Institute Amsterdam, 2Department of Anatomy, Embryology & Physiology, Academic Medical Center Amsterdam Here, we present a new technique to culture chick embryos ex ovo for time-lapse imaging using transmitted light and fluorescence microscopy. The submerged filter paper sandwich is a variant of the well-established filter paper carrier technique1 that allows for the culturing of chick embryos fully-submerged in a liquid culture medium. Bioengineering Real Time Monitoring of Intracellular Bile Acid Dynamics Using a Genetically Encoded FRET-based Bile Acid Sensor Sandra Van de Wiel1, Maarten Merkx2,3, Stan Van de Graaf1 1Tytgat Institute for Liver and Intestinal Research, Department of Gastroenterology and Hepatology, Academic Medical Center, 2Laboratory of Chemical Biology, Institute of Complex Molecular Systems (ICMS), 3Department of Biomedical Engineering, Eindhoven University of Technology We provide a detailed protocol to study bile acid dynamics in living cells using a genetically encoded BAS FRET sensor. This Bile Acid Sensor represents a unique tool to study (regulation of) bile acid transport and FXR activation in a wide range of cell types. Immunology and Infection Metabolic Characterization of Polarized M1 and M2 Bone Marrow-derived Macrophages Using Real-time Extracellular Flux Analysis Jan Van den Bossche1, Jeroen Baardman1, Menno P.J. de Winther1 1Department of Medical Biochemistry, Experimental Vascular Biology, Academic Medical Center, Amsterdam Metabolic reprogramming is a characteristic and prerequisite for M1 and M2 macrophage polarization. This manuscript describes an assay for the measurement of fundamental parameters of glycolysis and mitochondrial function in mouse bone marrow-derived macrophages. This tool can be applied to investigate how particular factors affect the macrophage’s metabolism and phenotype. Biology A Protocol for Lentiviral Transduction and Downstream Analysis of Intestinal Organoids Jooske F. Van Lidth de Jeude1, Jacqueline L. M. Vermeulen1, Paula S. Montenegro-Miranda1, Gijs R. Van den Brink1, Jarom Heijmans1 1Tytgat Institute for Liver and Intestinal Research, Department of Gasteroenterology and Hepatology, Academical Medical Center In this video protocol we give a step by step explanation of lentiviral transduction in organoids of primary intestinal epithelium and of processing and downstream analysis of these cultures by quantitative RT-PCR, RNA-microarray and immunohistochemistry. Medicine Human Dupuytren's Ex Vivo Culture for the Study of Myofibroblasts and Extracellular Matrix Interactions Sofia Karkampouna1, Peter Kloen2, Miryam C. Obdeijn3, Scott M. Riester4, Andre J. van Wijnen4,5, Marianna Kruithof-de Julio1,6 1Department of Molecular Cell Biology, Cancer Genomics Centre and Centre for Biomedical Genetics, Leiden University Medical Center, 2Department of Orthopedic Surgery, Academic Medical Center, 3Department of Plastic, Reconstructive and Hand Surgery, Academic Medical Center, 4Department of Orthopedic Surgery, Mayo Clinic, 5Department of Biochemistry and Molecular Biology, Mayo Clinic, 6Department of Dermatology, Leiden University Medical Center Dupuytren’s disease (DD) is a fibroproliferative disease of the palm of the hand. Here, we present a protocol to culture resection specimens from DD in a three-dimensional (3D) culture system. Such short-term culture system allows preservation of the 3D structure and molecular properties of the fibrotic tissue. Medicine In Vivo, Percutaneous, Needle Based, Optical Coherence Tomography of Renal Masses Peter G. Wagstaff1, Abel Swaan2, Alexandre Ingels1, Patricia J. Zondervan1, Otto M. van Delden3, Dirk J. Faber2, Ton G. van Leeuwen2, Jean J. de la Rosette1, Daniel M. de Bruin1,2, M. Pilar Laguna Pes1 1Department of Urology, Academic Medical Center, 2Department of Biomedical Engineering and Physics, Academic Medical Center, 3Department of Radiology, Academic Medical Center Optical coherence tomography (OCT) is a high resolution imaging technique that allows analysis of tissue specific optical properties providing the means for tissue differentiation. We developed needle based OCT, providing real-time imaging combined with on-the-spot tumor differentiation. This publication describes a method for percutaneous, needle based OCT of renal masses. Medicine Measuring Cardiac Autonomic Nervous System (ANS) Activity in Children Aimée E. van Dijk*1,2, René van Lien*3,4, Manon van Eijsden2,5, Reinoud J. B. J. Gemke6, Tanja G. M. Vrijkotte1, Eco J. de Geus3,4 1Department of Public Health, Academic Medical Center - University of Amsterdam, 2Department of Epidemiology, Documentation and Health Promotion, Public Health Service of Amsterdam (GGD), 3Department of Biological Psychology, VU University, 4EMGO+ Institute, VU University Medical Center, 5Institute of Health Sciences, VU University, 6Department of Pediatrics, VU University Medical Center Measurement of autonomic nervous system activity usually confines the researcher and participant to the laboratory, which may provide an intimidating environment to children. The VU University Ambulatory Monitoring System (VU-AMS) device can record cardiac autonomic control in any setting. The VU-AMS proved very amenable to testing in children. Medicine The Use of Pharmacological-challenge fMRI in Pre-clinical Research: Application to the 5-HT System Anne Klomp1, Jordi L. Tremoleda2, Anouk Schrantee1, Willy Gsell2, Liesbeth Reneman1 1Department of Radiology, Brain Imaging Center, Academic Medical Center Amsterdam, 2Biological Imaging Centre, MRC Clinical Sciences Centre, Imperial College London The goal of this technique is to assess serotonin (5-HT) neurotransmitter function in the live and free-breathing animal with pharmacological magnetic resonance imaging (phMRI) and an intravenous challenge with a selective serotonin reuptake inhibitor (SSRI), fluoxetine.