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Scuola Normale Superiore

6 ARTICLES PUBLISHED IN JoVE

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JoVE Journal

Fabrication, Operation and Flow Visualization in Surface-acoustic-wave-driven Acoustic-counterflow Microfluidics
Marco Travagliati 1,2, Richie Shilton 1, Fabio Beltram 2, Marco Cecchini 2
1NEST Center for Nanotechnology Innovation, Istituto Italiano di Tecnologia, 2NEST, Scuola Normale Superiore and Istituto Nanoscienze-CNR

In this video we first describe fabrication and operation procedures of a surface acoustic wave (SAW) acoustic counterflow device. We then demonstrate an experimental setup that allows for both qualitative flow visualization and quantitative analysis of complex flows within the SAW pumping device.

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Education

Synthesis, Cellular Delivery and In vivo Application of Dendrimer-based pH Sensors
Lorenzo Albertazzi 1, Barbara Storti 2, Marco Brondi 2, Sebastian Sulis Sato 2, Gian Michele Ratto 3, Giovanni Signore 4, Fabio Beltram 2
1Institute for Complex Molecular Systems & Laboratory of Macromolecular and Organic Chemistry, Eindhoven University of Technology & NEST, Scuola Normale Superiore and Istituto Nanoscienze-CNR, 2NEST, Scuola Normale Superiore and Istituto Nanoscienze-CNR & IIT@NEST, Center for Nanotechnology Innovation, 3NEST, Scuola Normale Superiore and Istituto Nanoscienze-CNR, 4IIT@NEST, Center for Nanotechnology Innovation

Fluorescence sensors are powerful tools in life science. Here we describe a methodology to synthesize and use dendrimer-based fluorescent sensors to measure pH in living cells and in vivo. The dendritic scaffold enhances the properties of conjugated fluorescent dyes leading to improved sensing properties.

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Bioengineering

From Fast Fluorescence Imaging to Molecular Diffusion Law on Live Cell Membranes in a Commercial Microscope
Carmine Di Rienzo 1,2, Enrico Gratton 3, Fabio Beltram 1, Francesco Cardarelli 2
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.

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Neuroscience

Exogenous Administration of Microsomes-associated Alpha-synuclein Aggregates to Primary Neurons As a Powerful Cell Model of Fibrils Formation
Giulia Panattoni 1,2, Lucia Rota 1, Emanuela Colla 1
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.

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Genetics

Modulation of Tau Subcellular Localization as a Tool to Investigate the Expression of Disease-related Genes
Giacomo Siano 1, Maria Claudia Caiazza 1, Martina Varisco 1, Mariantonietta Calvello 1, Valentina Quercioli 1, Antonino Cattaneo 1, Cristina Di Primio 1
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.

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Biology

Probing Structural and Dynamic Properties of Trafficking Subcellular Nanostructures by Spatiotemporal Fluctuation Spectroscopy
Gianmarco Ferri 1, Fabio Azzarello 1, Francesca D'Autilia 2, Francesco Cardarelli 1
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.

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