Clinical Academic Center of Coimbra 3 articles published in JoVE Biochemistry Analysis of Fluorescent-Stained Lipid Droplets with 3D Reconstruction for Hepatic Steatosis Assessment Karina Garcia1,2,3,4, André Alves1,2,3, Teresa M. Ribeiro-Rodrigues1,2,3, Flávio Reis1,2,3, Sofia Viana1,2,3,4 1Institute of Pharmacology & Experimental Therapeutics & Coimbra Institute for Clinical and Biomedical Research (iCBR), Faculty of Medicine, University of Coimbra, 2Center for Innovative Biomedicine and Biotechnology (CIBB), University of Coimbra, 3Clinical Academic Center of Coimbra (CACC), 4Polytechnic Institute of Coimbra, Coimbra Health School Herein, we demonstrate an optimized BODIPY 493/503 fluorescence-based protocol for lipid droplet characterization in liver tissue. Through the use of orthogonal projections and 3D reconstructions, the fluorophore allows for successful discrimination between microvesicular and macrovesicular steatosis and may represent a complementary approach to the classical histological protocols for hepatic steatosis assessment. Medicine Accessing the Cytotoxicity and Cell Response to Biomaterials Anabela Baptista Paula*1,2,3,4, Mafalda Laranjo*2,3,4,5, Ana Sofia Coelho1,2,3,4, Ana Margarida Abrantes2,3,4,5, Ana Cristina Gonçalves2,3,4,6, Ana Bela Sarmento-Ribeiro2,3,4,6, Manuel Marques Ferreira2,3,4,7, Maria Filomena Botelho2,3,4,5, Carlos Miguel Marto1,2,3,4,8, Eunice Carrilho1,2,3,4 1Institute of Integrated Clinical Practice, Faculty of Medicine, University of Coimbra, Coimbra, Portugal, 2Coimbra Institute for Clinical and Biomedical Research (iCBR) area of Environment Genetics and Oncobiology (CIMAGO), University of Coimbra, Coimbra, Portugal, 3Center for Innovative Biomedicine and Biotechnology (CIBB), University of Coimbra, Coimbra, Portugal, 4Clinical Academic Center of Coimbra, CACC, Portugal, 5Institute of Biophysics, Faculty of Medicine, University of Coimbra, Coimbra, Portugal, 6Laboratory of Oncobiology and Hematology, Faculty of Medicine, University of Coimbra, Coimbra, Portugal, 7Institute of Endodontics, Faculty of Medicine, University of Coimbra, Coimbra, Portugal, 8Institute of Experimental Pathology, Faculty of Medicine, University of Coimbra, Coimbra, Portugal This methodology aims to evaluate biomaterial cytotoxicity through the preparation of soluble extracts, using viability assays and phenotypic analysis, including flow cytometry, RT-PCR, immunocytochemistry, and other cellular and molecular biology techniques. Cancer Research Obtaining Cancer Stem Cell Spheres from Gynecological and Breast Cancer Tumors Mafalda Laranjo*1,2,3,4, Maria João Carvalho*1,2,3,4,5,6, Beatriz Serambeque1,2,3, André Alves2,7, Carlos Miguel Marto1,2,3,4,8, Isabel Silva9, Artur Paiva3,9,10, Maria Filomena Botelho1,2,3,4,8 1Institute of Biophysics, Faculty of Medicine, University of Coimbra, 2Coimbra Institute for Clinical and Biomedical Research (iCBR), area of Environment Genetics and Oncobiology (CIMAGO), Faculty of Medicine, University of Coimbra, 3CNC.IBILI/Center for Innovative Biomedicine and Biotechnology (CIBB), University of Coimbra, 4Clinical Academic Center of Coimbra (CACC), 5Universitary Clinic of Gynecology, Faculty of Medicine, University of Coimbra, 6Gynecology A Service, Coimbra Hospital and Universitary Center, 7Institute of Pharmacology & Experimental Therapeutics, Faculty of Medicine, University of Coimbra, 8Institute of Experimental Pathology, Faculty of Medicine, University of Coimbra, 9Cytometry Operational Management Unit, Clinical Pathology Service, Coimbra Hospital and Universitary Center, 10Polytechnic Institute of Coimbra, ESTESC-Coimbra Health School, Laboratory Biomedical Sciences The aim of this methodology is to identify cancer stem cells (CSC) in cancer cell lines and primary human tumor samples with the sphere-forming protocol, in a robust manner, using functional assays and phenotypic characterization with flow cytometry and Western blot.