The Barcelona Institute of Science and Technology (BIST) 7 articles published in JoVE Medicine Non-Invasive Monitoring of Microvascular Oxygenation and Reactive Hyperemia using Hybrid, Near-Infrared Diffuse Optical Spectroscopy for Critical Care M. Atif Yaqub*1, Marta Zanoletti*1, Lorenzo Cortese1, Daniel Senciales Sánchez1, Caterina Amendola2, Lorenzo Frabasile2, Umut Karadeniz1, Jacqueline Martinez Garcia1, Marta Martin1, Jordi Cortes-Picas1, Alba Caballer3, Edgar Cortes3, Sara Nogales3, Alberto Tosi4, Talyta Carteano5, Diego Sanoja Garcia5, Jakub Tomanik6, Tessa Wagenaar6, Hsiao Mui6, Claudia Nunzia Guadagno7, Shahrzad Parsa8, Sanathana Konugolu Venkata Sekar7, Luc Demarteau6, Tijl Houtbeckers6, Udo M. Weigel8, Michele Lacerenza9, Mauro Buttafava9, Alessandro Torricelli2,10, Davide Contini2, Jaume Mesquida3, Turgut Durduran1,11 1ICFO - Institut de Ciències Fotòniques, The Barcelona Institute of Science and Technology, 2Dipartimento di Fisica, Politecnico di Milano, 3Critical Care Department, Parc Taulí Hospital Universitari, 4Dipartimento di Elettronica, Informazione e Bioingegneria, Politecnico di Milano, 5ASPHALION s.l., 6Splendo, 7BioPixS-Biophotonics Standards, IPIC, Tyndall National Institute, 8Hemophotonics s.l., 9PIONIRS s.r.l., 10Consiglio Nazionale delle Ricerche, Istituto di Fotonica e Nanotecnologie, 11Institució Catalana de Recerca i Estudis Avançats (ICREA) We describe a protocol to non-invasively and continuously measure absolute microvascular blood flow index and blood oxygen saturation using a multi-modal device based on near-infrared diffuse optics. We then evaluate the metabolic rate of oxygen consumption and reactive hyperemia utilizing a vascular occlusion test. Biology High-Throughput Behavioral Aging and Lifespan Assays Using the Lifespan Machine Andrea Del Carmen-Fabregat1,2, Lucia Sedlackova1, Natasha Oswal1,2, Nicholas Stroustrup1,2 1Centre for Genomic Regulation (CRG), The Barcelona Institute of Science and Technology (BIST), 2Universitat Pompeu Fabra (UPF) The imaging platform "The Lifespan Machine" automates the lifelong observation of large populations. We show the steps required to perform lifespan, stress resistance, pathogenesis, and behavioral aging assays. The quality and scope of the data allow researchers to study interventions in aging despite the presence of biological and environmental variation. Neuroscience Calcium Imaging In Electrically Stimulated Flat-Mounted Retinas Marina Cunquero1, Maria Marsal1, Gustavo Castro-Olvera1, Steven T. Walston2, F. Taygun Duvan2, José Gabriel Macias-Montero3, Carles Puigdengoles3, Mokhtar Chmeissani3, Jose A. Garrido2,4, Pablo Loza-Alvarez1 1Institut de Ciències Fotòniques (ICFO), The Barcelona Institute of Science and Technology, 2Catalan Institute of Nanoscience and Nanotechnology (ICN2), CSIC and BIST, Campus UAB, 3Institut de Física d'Altes Energies (IFAE), The Barcelona Institute of Science and Technology, Campus UAB, 4Institució Catalana de Recerca i Estudis Avançats (ICREA) Retinal prostheses have the ability to generate visual perceptions. To advance the development of new prostheses, ex vivo methods are needed to test devices before implantation. This article provides a comprehensive protocol for studying calcium activity in the retinal ganglion cell layer when subjected to electrical stimulation. Neuroscience Hippocampal Neuronal Cultures to Detect and Study New Pathogenic Antibodies Involved in Autoimmune Encephalitis Marina Cunquero1, Esther Aguilar2, Pablo Loza-Alvarez1, Jesús Planagumà1,2 1ICFO-Institut de Ciències Fotòniques, The Barcelona Institute of Science and Technology, 2Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Hospital Clínic, Universitat de Barcelona Autoimmune encephalitis is a new category of antibody-mediated diseases of the central nervous system. Hippocampal neurons can be used to discover and characterize these antibodies. This article provides a protocol for primary cell culture and immunostaining to determine autoantibodies in the serum and cerebrospinal fluid of patients. Developmental Biology Direct Force Measurements of Subcellular Mechanics in Confinement using Optical Tweezers Frederic Català-Castro1, Valeria Venturini2,3, Santiago Ortiz-Vásquez1, Verena Ruprecht2,4, Michael Krieg1 1Neurophotonics and Mechanical Systems Biology, Institut de Ciències Fotòniques, ICFO, 2Center for Genomic Regulation (CRG), The Barcelona Institute of Science and Technology, 3Institut de Ciències Fotòniques, ICFO, 4Universitat Pompeu Fabra (UPF) Here, we present a protocol to investigate the intracellular mechanical properties of isolated embryonic zebrafish cells in three-dimensional confinement with direct force measurement by an optical trap. Genetics Identification of Enhancer-Promoter Contacts in Embryoid Bodies by Quantitative Chromosome Conformation Capture (4C) Tian V. Tian1,2,3, Enrique Vidal1,2, Thomas Graf1,2, Grégoire Stik1,2 1Center for Genomic Regulation (CRG), The Barcelona Institute of Science and Technology, 2Universitat Pompeu Fabra, 3 We report the application of quantitative chromosome conformation capture followed by high-throughput sequencing in embryoid bodies generated from embryonic stem cells. This technique allows to identify and quantitate the contacts between putative enhancers and promoter regions of a given gene during embryonic stem cell differentiation. Bioengineering Dendrimer-based Uneven Nanopatterns to Locally Control Surface Adhesiveness: A Method to Direct Chondrogenic Differentiation Ignasi Casanellas1,2, Anna Lagunas3,1, Iro Tsintzou1, Yolanda Vida4,5, Daniel Collado4,5, Ezequiel Pérez-Inestrosa4,5, Cristina Rodríguez-Pereira6, Joana Magalhaes3,6, Pau Gorostiza1,3,7, José A. Andrades8,3, José Becerra8,3,5, Josep Samitier1,3,2 1Institute for Bioengineering of Catalonia (IBEC), The Barcelona Institute of Science and Technology (BIST), 2Department of Engineering Electronics, University of Barcelona (UB), 3Networking Biomedical Research Center (CIBER), 4Instituto de Investigacin Biomédica de Málaga (IBIMA), Department of Organic Chemistry, Universidad de Málaga (UMA), 5Andalusian Centre for Nanomedicine and Biotechnology-BIONAND, 6Unidad de Bioingeniería Tisular y Terapia Celular (GBTTC-CHUAC), Grupo de Reumatolog ía, Instituto de Investigación Biomèdica de A Coruña (INIBIC), Complexo Hospitalario Universitario de A Coruña (CHUAC), Sergas, Universidade da Coruña (UDC), 7Institució Catalana de Recerca i Estudis Avançats (ICREA), 8Instituto de Investigación Biomédica de Málaga (IBIMA), Department of Cell Biology, Genetics and Physiology, Universidad de Málaga (UMA) A method to obtain dendrimer-based uneven nanopatterns that permit the nanoscale control of local arginine-glycine-aspartic acid (RGD) surface density is described and applied for the study of cell adhesion and chondrogenic differentiation.