Universita della Svizzera italiana 3 articles published in JoVE Neuroscience Targeting Alpha Synuclein Aggregates in Cutaneous Peripheral Nerve Fibers by Free-floating Immunofluorescence Assay Elena Vacchi1, Sandra Pinton1, Alain Kaelin-Lang1,2,3,4, Giorgia Melli1,2 1Laboratory for Biomedical Neurosciences, Neurocenter of Southern Switzerland, 2Neurology Department, Neurocenter of Southern Switzerland, 3Department of Neurology, Inselspital, Bern University Hospital, University of Bern, 4Faculty of Biomedical Sciences, Università della Svizzera Italiana Here, we present a protocol for a free-floating indirect immunofluorescence assay on skin biopsy sections that allows for the identification of disease specific conformation variants of alpha synuclein involved in Parkinson disease and multiple proteins of the peripheral nervous system. Biology Flow Cytometric Analysis of Extracellular Vesicles from Cell-conditioned Media Carolina Balbi*1, Sara Bolis*1, Giuseppe Vassalli1,2,3, Lucio Barile1 1Cellular and Molecular Cardiology Laboratory, Cardiocentro Ticino Foundation, Switzerland, 2Molecular Cardiology Institute, Dept. of Cardiology, University of Zürich, 3Faculty of Biomedical Science, Università Svizzera Italiana, Switzerland The protocol describes a reproducible method designed for use with cell culture supernatants to detect surface epitopes on small extracellular vesicles (EV). It utilizes specific EV immunoprecipitation using beads coupled with antibodies that recognize surface antigen CD9, CD63, and CD81. The method is optimized for downstream flow cytometry analysis. Immunology and Infection Imaging Cell Interaction in Tracheal Mucosa During Influenza Virus Infection Using Two-photon Intravital Microscopy Miguel Palomino-Segura*1,2, Tommaso Virgilio*1,2, Diego Morone1, Diego U. Pizzagalli1,3, Santiago F. Gonzalez1 1Faculty of Biomedical Sciences, Institute for Research in Biomedicine, Università della Svizzera italiana (USI), 2Graduate School of Cellular and Molecular Sciences, Faculty of Medicine, University of Bern, 3Institute of Computational Science, Università della Svizzera italiana (USI) In this study, we present a protocol to perform two-photon intravital imaging and cell interaction analysis in the murine tracheal mucosa after infection with influenza virus. This protocol will be relevant for researchers studying immune cell dynamics during respiratory infections.