Politecnico di Milano View Institution's Website 13 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. Developmental Biology AMEBaS: Automatic Midline Extraction and Background Subtraction of Ratiometric Fluorescence Time-Lapses of Polarized Single Cells Rafael Badain1, Daniel S. C. Damineli1,2, Maria Teresa Portes3, José Feijó4, Stefano Buratti5, Giorgia Tortora6, Hugo Neves de Oliveira1, Roberto M. Cesar Jr1 1Institute of Mathematics and Statistics, University of São Paulo, 2Center for Mathematics, Computing and Cognition, Federal University of ABC, 3Department of Botany, Institute of Biosciences, University of São Paulo, 4Cell Biology and Molecular Genetics Department, University of Maryland, 5Department of Biosciences, University of Milan, 6Department of Physics, Politecnico di Milano Current methods for analyzing the intracellular dynamics of polarized single cells are often manual and lack standardization. This manuscript introduces a novel image analysis pipeline for automating midline extraction of single polarized cells and quantifying spatiotemporal behavior from time lapses in a user-friendly online interface. Engineering Multiplex Chemical Imaging Based on Broadband Stimulated Raman Scattering Microscopy Alejandro De la Cadena1, Federico Vernuccio1, Benedetta Talone1, Arianna Bresci1, Chiara Ceconello1, Subir Das1, Renzo Vanna2, Giulio Cerullo1,2, Dario Polli1,2 1Dipartimento di Fisica, Politecnico di Milano, 2Institute for Photonics and Nanotechnologies (IFN-CNR) We present a protocol to acquire chemical images with broadband stimulated Raman scattering (SRS) microscopy. Based on an SRS microscope that operates with differential multichannel-lock-in detection, the protocol describes the sample preparation, adjustment of the SRS apparatus, and chemometrics to disentangle different constituents of chemically heterogeneous samples. Medicine International Expert Consensus and Recommendations for Neonatal Pneumothorax Ultrasound Diagnosis and Ultrasound-guided Thoracentesis Procedure Jing Liu1,2, Dalibor Kurepa3, Francesco Feletti4,5, Almudena Alonso-Ojembarrena6, Jovan Lovrenski7, Roberto Copetti8, Erich Sorantin9, Javier Rodriguez-Fanjul10, Karishma Katti3, Andrea Aliverti4, Huayan Zhang11,12, Misun Hwang13, Tsu F. Yeh14, Cai-Bao Hu15, Xing Feng16, Ru-Xin Qiu1,2, Jing-Han Chi17, Li-Li Shang18, Guo-Rong Lyu19, Shao-Zheng He20, Yan-Fen Chai21, Zhan-Jun Qiu22, Hai-Ying Cao2,23, Yue-Qiao Gao1,2, Xiao-Ling Ren1,2, Guo Guo1,24, Li Zhang1,2, Ying Liu1,2, Wei Fu1,2, Zu-Lin Lu1,2, Hong-Lei Li1,2 1Department of Neonatology and NICU, Beijing Chaoyang District Maternal and Child Healthcare Hospital, 2The National Neonatal Lung Ultrasound Training Base, 3 Pneumothorax is a common emergency and critical disease in newborn infants that needs rapid, clear diagnosis and timely treatment. Diagnosis and treatment based on chest X-rays are associated with delayed management and radiation damage. Lung ultrasound (US) provides useful guidance for rapid, accurate diagnosis and the precise thoracentesis of pneumothorax. Biology Exploiting Live Imaging to Track Nuclei During Myoblast Differentiation and Fusion Giorgia Careccia*1,2, Federica Colombo*1,3, Mario Tirone1, Alessandra Agresti1, Marco E. Bianchi1,2, Samuel Zambrano*1,2, Emilie Vénéreau*1 1Chromatin Dynamics Unit, Istituto di Ricovero e Cura a Carattere Scientifico (IRCCS) San Raffaele Scientific Institute, 2Vita-Salute San Raffaele University, 3Department of Electronics, Information and Bioengineering, Politecnico di Milano Skeletal muscle differentiation is a highly dynamic process, which particularly relies on nuclear positioning. Here, we describe a method to track nuclei movements by live cell imaging during myoblast differentiation and myotube formation and to perform a quantitative characterization of nuclei dynamics by extracting information from automatic tracking. Chemistry From Molecules to Materials: Engineering New Ionic Liquid Crystals Through Halogen Bonding Gabriella Cavallo1, Duncan W. Bruce2, Giancarlo Terraneo1, Giuseppe Resnati3, Pierangelo Metrangolo1 1Laboratory of Supramolecular and Bio-Nanomaterials (SBNLab), Department of Chemistry, Materials, and Chemical Engineering "Giulio Natta", Politecnico di Milano, 2Department of Chemistry, University of York, 3Laboratory of Nanostructured Fluorinated Materials (NFMLab), Department of Chemistry, Materials, and Chemical Engineering "Giulio Natta", Politecnico di Milano A protocol for the synthesis of a new type of mesogens, based on the halogen-bonded supramolecular anion [CnF2n+1-I···I···I-CnF2n+1]−, is reported. Chemistry Laser-heating and Radiance Spectrometry for the Study of Nuclear Materials in Conditions Simulating a Nuclear Power Plant Accident Dario Manara1, Luca Soldi1,2,4, Sara Mastromarino1,3,5, Kostantinos Boboridis1, Davide Robba1, Luka Vlahovic1, Rudy Konings1 1European Commission, Joint Research Centre, 2Energy Department, Politecnico di Milano, 3Department of Chemical Physics, Sapienza - Università di Roma, 4CEA Saclay, 5TU Delft We present experiments in which real nuclear fuel, cladding, and containment materials are laser heated to temperatures beyond 3,000 K while their behavior is studied by radiance spectroscopy and thermal analysis. These experiments simulate, on a laboratory scale, the formation of a lava-phase following a nuclear reactor core meltdown. Environment Image-based Lagrangian Particle Tracking in Bed-load Experiments Alessio Radice1, Sankar Sarkar2, Francesco Ballio1 1Dept. of Civil and Environmental Engineering, Politecnico di Milano, 2Physics and Applied Mathematics Unit, Indian Statistical Institute The manuscript presents a protocol for the conduction of bed-load sediment transport experiments where the moving particles are tracked by image analysis. The experimental facility, the procedures for run realization and data processing, and finally some proof-of-concept results are presented here. Bioengineering The Synthesis of RGD-functionalized Hydrogels as a Tool for Therapeutic Applications Emanuele Mauri1, Alessandro Sacchetti1, Filippo Rossi1 1Department of Chemistry, Materials and Chemical Engineering "Giulio Natta", Politecnico di Milano We present a protocol for the synthesis of RGD-functionalized hydrogels as devices for cell and drug delivery. The procedure involves copper catalyzed alkyne-azide cycloaddition (CuAAC) between alkyne-modified polyacrylic acid (PAA) and a RGD-azide derivative. The hydrogels are formed using microwave-assisted polycondensation and their physicochemical properties are investigated. Bioengineering Transport Properties of Ibuprofen Encapsulated in Cyclodextrin Nanosponge Hydrogels: A Proton HR-MAS NMR Spectroscopy Study Monica Ferro*1, Franca Castiglione*1, Carlo Punta1, Lucio Melone1,2, Walter Panzeri3, Barbara Rossi4, Francesco Trotta5, Andrea Mele1 1Department of Chemistry, Materials and Chemical Engineering "G. Natta", Politecnico di Milano, 2Università degli studi e-Campus, 3ICRM, CNR - Consiglio Nazionale delle Ricerche, 4Elettra Sincrotrone Trieste, 5Department of Chemistry, University of Torino The motion regimes of ibuprofen encapsulated in β-cyclodextrin nanosponges polymer network are investigated using pulsed-field-gradient spin-echo (PGSE) NMR technique. Synthesis, purification, drug loading, implementation of the NMR pulse sequence and data analysis to work out the mean square displacement of the drug at several observation times are described in detail. Bioengineering Optical Control of Living Cells Electrical Activity by Conjugated Polymers Nicola Martino1, Caterina Bossio1, Susana Vaquero Morata1, Guglielmo Lanzani1,2, Maria Rosa Antognazza1 1Center for Nano Science and Technology @PoliMi, Istituto Italiano di Tecnologia, 2Dipartimento di Fisica, Politecnico di Milano A method for stimulation of in-vitro cell cultures electrical activity with visible light, based on the use of organic semiconducting polymers is described. Bioengineering Engineering 3D Cellularized Collagen Gels for Vascular Tissue Regeneration Sébastien Meghezi1, Dawit G. Seifu1,2, Nina Bono1,3, Larry Unsworth2,4,5, Kibret Mequanint6, Diego Mantovani1,2 1Laboratory for Biomaterials and Bioengineering, Department Min-Met-Materials Eng & CHU de Québec Research Center, Canada Research Chair I for the Innovation in Surgery, Laval University, 2NSERC CREATE Program for Regenerative Medicine (NCPRM), Laval University, 3Department Electronics, Information and Bioengineering, Politecnico di Milano, 4Department of Chemical and Materials Engineering, University of Alberta, 5National Institute for Nanotechnology, National Research Council (Canada), 6Department of Chemical and Biochemical Engineering, University of Western Ontario In this work, we present a technique for the rapid fabrication of living vascular tissues by direct culturing of collagen, smooth muscle cells and endothelial cells. In addition, a new protocol for the mechanical characterization of engineered vascular tissues is described. Engineering Fabrication of Nano-engineered Transparent Conducting Oxides by Pulsed Laser Deposition Paolo Gondoni1, Matteo Ghidelli1, Fabio Di Fonzo1,2, Andrea Li Bassi1,2, Carlo S. Casari1 1Department of Energy and NEMAS - Center for NanoEngineered Materials and Surfaces, Politecnico di Milano, 2Center for Nano Science and Technology, Instituto Italiano di Tecnologia We describe the experimental method to deposit nanostructured oxide thin films by nanosecond Pulsed Laser Deposition (PLD) in the presence of a background gas. By using this method Al-doped ZnO (AZO) films, from compact to hierarchically structured as nano-tree forests, can be deposited.
