Universidade de Lisboa 11 articles published in JoVE Engineering Membraneless Hydrogen Peroxide Fuel Cells as a Promising Clean Energy Source Fenyang Zhu1, Guoxiang Chen1, Aleksei Kuzin1,2, Dmitry A. Gorin2, Brij Mohan3, Gaoshan Huang1, Yongfeng Mei1,4,5,6,7, Alexander A. Solovev1 1Department of Materials Science & State Key Laboratory of Molecular Engineering of Polymers, Fudan University, 2Center for Photonic Science and Engineering, Skolkovo Institute of Science and Technology, 3Centro de Quimica Estrutural, Institute of Molecular Sciences, Instituto Superior Tecnico, Universidade de Lisboa, 4Center for Biomedical Engineering, School of Information Science and Technology, Fudan University, 5International Institute of Intelligent Nanorobots and Nanosystems, Fudan University, 6Shanghai Frontiers Science Research Base of Intelligent Optoelectronics and Perception, Fudan University, 7Yiwu Research Institute of Fudan University This protocol introduces the design and evaluation of innovative three-dimensional electrodes for hydrogen peroxide fuel cells, utilizing Au-electroplated carbon fiber cloth and Ni-foam electrodes. The research findings highlight hydrogen peroxide's potential as a promising candidate for sustainable energy technologies. Medicine Micro-Colony Forming Unit Assay for Efficacy Evaluation of Vaccines Against Tuberculosis David Pires1,2,3, Paulo J. G. Bettencourt1,2 1Faculty of Medicine, Universidade Católica Portuguesa, 2Center for Interdisciplinary Research in Health, Universidade Católica Portuguesa, 3Host-Pathogen Interactions Unit, Research Institute for Medicines, iMed-ULisboa, Faculty of Pharmacy, Universidade de Lisboa The determination of colony-forming units (CFU) is the gold-standard technique for quantifying bacteria, including Mycobacterium tuberculosis which can take weeks to form visible colonies. Here we describe a micro-CFU for CFU determination with increased time efficiency, reduced lab space and reagent cost, and scalability to medium and high throughput experiments. Chemistry Dose Uptake of Platinum- and Ruthenium-based Compound Exposure in Zebrafish by Inductively Coupled Plasma Mass Spectrometry with Broader Applications Brittany F. Karas1,2, Cathleen L. Doherty1, Kristin R. Terez2, Leonor Côrte-Real3, Keith R. Cooper1,2, Brian T. Buckley1 1Environmental and Occupational Health Sciences Institute, Rutgers University, 2Department of Biochemistry and Microbiology, Rutgers University, 3Centro de Química Estrutural and Departamento de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa The increased rate of pharmaco- and toxicokinetic analyses of metals and metal-based compounds in zebrafish can be advantageous for environmental and clinical translation studies. The limitation of unknown waterborne exposure uptake was overcome by conducting trace metal analysis on digested zebrafish tissue using inductively coupled plasma mass spectrometry. Bioengineering Scalable Generation of Mature Cerebellar Organoids from Human Pluripotent Stem Cells and Characterization by Immunostaining Teresa P. Silva1,2, Tiago G. Fernandes1, Diogo E. S. Nogueira1, Carlos A. V. Rodrigues1, Evguenia P. Bekman1,2,3, Yas Hashimura4, Sunghoon Jung4, Brian Lee4, Maria Carmo-Fonseca2, Joaquim M. S. Cabral1 1iBB - Institute for Bioengineering and Biosciences and Department of Bioengineering, Instituto Superior Técnico, Universidade de Lisboa, 2Instituto de Medicina Molecular João Lobo Antunes, Faculdade de Medicina, Universidade de Lisboa, 3The Discoveries Centre for Regenerative and Precision Medicine, Lisbon Campus, Universidade de Lisboa, 4PBS Biotech, Inc, Camarillo, CA, USA This protocol describes a dynamic culture system to produce controlled size aggregates of human pluripotent stem cells and further stimulate differentiation in cerebellar organoids under chemically-defined and feeder-free conditions using a single-use bioreactor. Neuroscience Isolation and Expansion of Neurospheres from Postnatal (P1−3) Mouse Neurogenic Niches Rita Soares1,2,3, Filipa F. Ribeiro1,2, Diogo M. Lourenço1,2, Rui S. Rodrigues1,2, João B. Moreira1,2, Ana M. Sebastião1,2, Vanessa A. Morais1,3, Sara Xapelli1,2 1Instituto de Medicina Molecular João Lobo Antunes, Faculdade de Medicina, Universidade de Lisboa, 2Instituto de Farmacologia e Neurociências, Faculdade de Medicina, Universidade de Lisboa, 3Instituto de Biologia Molecular, Faculdade de Medicina, Universidade de Lisboa In this article, we describe, in detail, a protocol for the generation of neurosphere cultures from postnatal mouse neural stem cells derived from the main mouse neurogenic niches. Neurospheres are used to identify neural stem cells from brain tissue allowing the estimation of precursor cell numbers. Moreover, these 3D structures can be plated in differentiative conditions, giving rise to neurons, oligodendrocytes and astrocytes, allowing the study of cell fate. Biochemistry A Rapidly Incremented Tethered-Swimming Maximal Protocol for Cardiorespiratory Assessment of Swimmers Dalton M. Pessôa Filho*1,2, Danilo A. Massini*2, Leandro O. C. Siqueira*2, Luiz Gustavo A. Santos*2, Camila M. T. Vasconcelos*2, Tiago A. F. Almeida*1,3, Mário A. C. Espada*4,5, Joana F. Reis*3,6, Francisco B. Alves*3, Fred J. DiMenna*7,8 1Department of Physical Education, São Paulo State University (UNESP) at Bauru, 2Institute of Bioscience, Graduate Program in Human Development and Technology, São Paulo State University (UNESP) at Rio Claro, 3Ciper, Faculdade de Motricidade Humana, Universidade de Lisboa, 4Department of Science and Technology, School of Education, Polytechnic Institute of Setúbal, 5Quality of Life Research Center, Polytechnic Institute of Santarem, 6Universidade Europeia at Lisbon, 7Division of Endocrinology, Diabetes and Bone Disease, Icahn School of Medicine at Mount Sinai, 8Department of Biobehavioral Sciences, Teachers College, Columbia University As opposed to measurement during free swimming, which presents inherent challenges and limitations, determination of important parameters of cardiorespiratory function for swimmers can be made using a more feasible and easier to administer tethered-swimming rapidly incremented protocol with gas exchange and ventilatory data collection. Medicine Echocardiographic Assessment of Cardiac Anatomy and Function in Adult Rats Sónia Ribeiro*1,2, Ana Rita S. Pereira*1, Ana Teresa Pinto1, Filipe Rocha1, Augusto Ministro1,2, Manuela Fiuza1,2, Fausto Pinto1,2, Susana Constantino Rosa Santos1 1Centro Cardiovascular da Universidade de Lisboa, Faculdade de Medicina, Universidade de Lisboa, 2Departamento Coração e Vasos, Centro Hospitalar Universitário Lisboa Norte A non-invasive protocol for transthoracic echocardiography assessment of cardiac anatomy and function for adult rats is presented in the current study. The heart valves, all four cardiac chambers and the ascending aorta, aortic arch and descending aorta are studied in detail. Immunology and Infection Detection of Extravascular Trypanosoma Parasites by Fine Needle Aspiration Tânia Carvalho1, Ana Margarida Santos1, Luísa M. Figueiredo2 1Comparative Pathology Unit, Instituto de Medicina Molecular – Faculdade de Medicina de Lisboa, 2Biology of Parasitism Laboratory, Instituto de Medicina Molecular – Faculdade de Medicina de Lisboa Fine Needle Aspiration is a technique, whereby cells are obtained from a lesion or organ using a thin needle. Aspirated material is smeared, stained and examined under a microscope for diagnosis or used for molecular biology, cytometry or in vitro analysis. It is cheap, simple, quick and causes minimal trauma. Developmental Biology Isolation of Embryonic Tissues and Formation of Quail-Chicken Chimeric Organs Using The Thymus Example Marta Figueiredo1, Hélia Neves1 1Instituto de Histologia e Biologia do Desenvolvimento, Faculdade de Medicina, Universidade de Lisboa This article provides a method to isolate pure embryonic tissues from quail and chicken embryos that can be combined to form ex vivo chimeric organs. Developmental Biology Two-step Approach to Explore Early- and Late-stages of Organ Formation in the Avian Model: The Thymus and Parathyroid Glands Organogenesis Paradigm Marta Figueiredo1, Hélia Neves1 1Instituto de Histologia e Biologia do Desenvolvimento, Faculdade de Medicina, Universidade de Lisboa This article provides an updated approach to the classical quail-chicken chimera system to study organ formation, by combining novel in vitro and in ovo experimental procedures. Developmental Biology In Vitro Differentiation of Mature Myofibers for Live Imaging Mafalda R. Pimentel1, Sestina Falcone2, Bruno Cadot2, Edgar R. Gomes1,2 1Instituto de Medicina Molecular, Faculdade de Medicina, Universidade de Lisboa, 2Myology Research Center, UM76-INSERM U974-CNRS FRE 361, Sorbonne University, UPMC University of Paris 6 Muscle cells are among the most complex eukaryotic cells. We present a protocol for the in vitro differentiation of highly mature myofibers that allows for genetic manipulation and clear imaging during all developmental stages.
