Scuola Normale Superiore 4 articles published in JoVE Biology Probing Structural and Dynamic Properties of Trafficking Subcellular Nanostructures by Spatiotemporal Fluctuation Spectroscopy Gianmarco Ferri1, Fabio Azzarello1, Francesca D'Autilia2, Francesco Cardarelli1 1NEST Laboratory, Scuola Normale Superiore, 2Center for Nanotechnology Innovation@NEST CNI@NEST Imaging-derived mean square displacement (iMSD) analysis is applied to macropinosomes to highlight their intrinsic time-evolving nature in terms of structural and dynamic properties. Macropinosomes are then compared to insulin secretory granules (ISGs) as a reference for subcellular structures with time-invariant average structural/dynamic properties. Genetics Modulation of Tau Subcellular Localization as a Tool to Investigate the Expression of Disease-related Genes Giacomo Siano1, Maria Claudia Caiazza1, Martina Varisco1, Mariantonietta Calvello1, Valentina Quercioli1, Antonino Cattaneo1, Cristina Di Primio1 1Laboratory of Biology, BIO@SNS, Scuola Normale Superiore Tau is a neuronal protein present both in the cytoplasm, where it binds microtubules, and in the nucleus, where it exerts unconventional functions including the modulation of Alzheimer's disease-related genes. Here, we describe a method to investigate the function of nuclear Tau while excluding any interferences coming from cytoplasmic Tau. Neuroscience Exogenous Administration of Microsomes-associated Alpha-synuclein Aggregates to Primary Neurons As a Powerful Cell Model of Fibrils Formation Giulia Panattoni1,2, Lucia Rota1, Emanuela Colla1 1Bio@SNS Laboratory, Scuola Normale Superiore, 2International School for Advanced Studies (SISSA/ISAS) The goal of this protocol is to provide a cell-based system that replicates the formation of alpha-synuclein aggregates in vivo. Intracellular alpha-synuclein inclusions are seeded in primary neurons by the internalization and propagation of exogenous administered native microsomes-associated alpha-synuclein aggregates isolated from diseased alpha-synuclein transgenic mice. Bioengineering From Fast Fluorescence Imaging to Molecular Diffusion Law on Live Cell Membranes in a Commercial Microscope Carmine Di Rienzo1,2, Enrico Gratton3, Fabio Beltram1, Francesco Cardarelli2 1NEST Laboratory, Scuola Normale Superiore, 2Center for Nanotechnology Innovation, Instituto Italiano di Tecnologia, 3Laboratory for Fluorescence Dynamics, Department of Biomedical Engineering, University of California, Irvine Spatial distribution and temporal dynamics of plasma membrane proteins and lipids is a hot topic in biology. Here this issue is addressed by a spatio-temporal image fluctuation analysis that provides conceptually the same physical quantities of single particle tracking, but it uses small molecular labels and standard microscopy setups.
