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École Polytechnique Fédérale de Lausanne

11 ARTICLES PUBLISHED IN JoVE

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Neuroscience

Focussed Ion Beam Milling and Scanning Electron Microscopy of Brain Tissue
Graham Knott 1, Stéphanie Rosset 1, Marco Cantoni 1
1Centre of interdisciplinary electron microscopy, École Polytechnique Fédérale de Lausanne

This protocol describes how resin embedded brain tissue can be prepared and imaged in the three dimensions in the focussed ion beam, scanning electron microscope.

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Medicine

How to Measure Cortical Folding from MR Images: a Step-by-Step Tutorial to Compute Local Gyrification Index
Marie Schaer 1, Meritxell Bach Cuadra 2,3, Nick Schmansky 4, Bruce Fischl 4, Jean-Philippe Thiran 2, Stephan Eliez 1
1Department of Psychiatry, University of Geneva School of Medicine, 2Signal Processing Laboratory, École Polytechnique Fédérale de Lausanne, 3Department of Radiology, University Hospital Center and University of Lausanne, 4Athinoula A. Martinos Center for Biomedical Imaging, Massachusetts General Hospital

Measuring gyrification (cortical folding) at any age represents a window into early brain development. Hence, we previously developed an algorithm to measure local gyrification at thousands of points over the hemisphere1. In this paper, we detail the computation of this local gyrification index.

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Engineering

Angle-resolved Photoemission Spectroscopy At Ultra-low Temperatures
Sergey V. Borisenko 1, Volodymyr B. Zabolotnyy 1, Alexander A. Kordyuk 1,2, Danil V. Evtushinsky 1, Timur K. Kim 1,3, Emanuela Carleschi 4, Bryan P. Doyle 4, Rosalba Fittipaldi 5, Mario Cuoco 5, Antonio Vecchione 5, Helmut Berger 6
1Institute for Solid State Research, IFW-Dresden, 2Institute of Metal Physics of National Academy of Sciences of Ukraine, 3Diamond Light Source LTD, 4Department of Physics, University of Johannesburg, 5CNR-SPIN, and Dipartimento di Fisica "E. R. Caianiello", Università di Salerno, 6Institute of Physics of Complex Matter, École Polytechnique Fédérale de Lausanne

The overall goal of this method is to determine the low-energy electronic structure of solids at ultra-low temperatures using Angle-Resolved Photoemission Spectroscopy with synchrotron radiation.

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Bioengineering

Harmonic Nanoparticles for Regenerative Research
Flavio Ronzoni 1, Thibaud Magouroux 2, Remi Vernet 1, Jérôme Extermann 3, Darragh Crotty 4, Adriele Prina-Mello 5, Daniel Ciepielewski 6, Yuri Volkov 4, Luigi Bonacina 2, Jean-Pierre Wolf 2, Marisa Jaconi 1
1Department of Pathology and Immunology, Faculty of Medicine, University of Geneva, 2Physics Department, GAP-Biophotonics, University of Geneva, 3Laboratoire d'Optique Biomédicale (LOB), Faculté des Sciences et Techniques de l'Ingénieur, École Polytechnique Fédérale de Lausanne, 4Department of Clinical Medicine, School of Medicine, Trinity College Dublin, 5School of Medicine and CRANN, Trinity College Dublin, 6Nikon AG Instruments

Protocol details are provided for in vitro labeling human embryonic stem cells with second harmonic generating nanoparticles. Methodologies for hESC investigation by multi-photon microscopy and their differentiation into cardiac clusters are also presented.

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Bioengineering

Long-term Intravital Immunofluorescence Imaging of Tissue Matrix Components with Epifluorescence and Two-photon Microscopy
Esra Güç *1, Manuel Fankhauser *1, Amanda W. Lund 1,2, Melody A. Swartz 1, Witold W. Kilarski 1
1Institute of Bioengineering and Swiss Institute of Experimental Cancer Research (ISREC), École Polytechnique Fédérale de Lausanne, 2Department of Cell and Developmental Biology and Knight Cancer Institute, Oregon Health & Science University

The extracellular matrix undergoes substantial remodeling during wound healing, inflammation and tumorigenesis. We present a novel intravital immunofluorescence microscopy approach to visualize the dynamics of fibrillar as well as mesh-like matrix components with high spatial and temporal resolution using epifluorescence or two-photon microscopy.

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Engineering

Monovalent Cation Doping of CH3NH3PbI3 for Efficient Perovskite Solar Cells
Mojtaba Abdi-Jalebi 1, M. Ibrahim Dar 2, Aditya Sadhanala 1, Satyaprasad P. Senanayak 1, Michael Grätzel 2, Richard H. Friend 1
1Cavendish Laboratory, University of Cambridge, 2Institute of Chemical Sciences and Engineering, École Polytechnique Fédérale de Lausanne

Here, we present a protocol to adjust the properties of solution-processed CH3NH3PbI3 through the incorporation of monovalent cation additives in order to achieve highly efficient perovskite solar cells.

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Biology

Quantifying Leukocyte Egress via Lymphatic Vessels from Murine Skin and Tumors
Maria M. Steele 1, Madeline J. Churchill 2, Alec P. Breazeale 1, Ryan S. Lane 1, Nicholas A. Nelson 1, Amanda W. Lund 1,2,3,4
1Department of Cell, Developmental, & Cancer Biology, Oregon Health and Science University, 2Department of Molecular Microbiology & Immunology, Oregon Health and Science University, 3Department of Dermatology, Oregon Health and Science University, 4Knight Cancer Institute, Oregon Health and Science University

Here, we demonstrate the methods for in vivo quantification of leukocyte egress from naïve, inflamed, and malignant murine skin. We perform a head-to-head comparison of two models: transdermal FITC application and in situ photoconversion. Furthermore, we demonstrate the utility of photoconversion for tracking leukocyte egress from cutaneous tumors.

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Chemistry

Synthesis and Characterization of Amphiphilic Gold Nanoparticles
Zekiye P. Guven *1, Paulo H. Jacob Silva *1, Zhi Luo 1, Urszula B. Cendrowska 1, Matteo Gasbarri 1, Samuel T. Jones 1,2, Francesco Stellacci 1,3
1Institute of Materials, École Polytechnique Fédérale de Lausanne, 2School of Materials, University of Manchester, 3Interfaculty Institute of Bioengineering, École Polytechnique Fédérale de Lausanne

Amphiphilic gold nanoparticles can be used in many biological applications. A protocol to synthesize gold nanoparticles coated by a binary mixture of ligands and a detailed characterization of these particles is presented.

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Environment

Automated 3D Optical Coherence Tomography to Elucidate Biofilm Morphogenesis Over Large Spatial Scales
Anna Depetris 1, Antoine Wiedmer 2, Michael Wagner 3, Sebastian Schäfer 4, Tom J. Battin 1, Hannes Peter 1
1Stream Biofilm and Ecosystem Research Laboratory, School of Architecture, Civil and Environmental Engineering, Ecole Polytechnique Fédérale de Lausanne, 2Plateforme technique, School of Architecture, Civil and Environmental Engineering, Ecole Polytechnique Fédérale de Lausanne, 3Water Chemistry and Water Technology, Engler-Bunte-Institut, Karlsruhe Institute of Technology, 4Thorlabs GmbH

Microbial biofilms form complex architectures at interphases and develop into highly scale-dependent spatial patterns. Here, we introduce an experimental system (hard- and software) for the automated acquisition of 3D optical coherence tomography (OCT) datasets. This toolset allows the non-invasive and multi-scale characterization of biofilm morphogenesis in space and time.

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Environment

Combining Fluidic Devices with Microscopy and Flow Cytometry to Study Microbial Transport in Porous Media Across Spatial Scales
David Scheidweiler 1,2, Pietro De Anna 2, Tom J. Battin 1, Hannes Peter 1
1Stream Biofilm and Ecosystem Research Laboratory, École Polytechnique Fédérale de Lausanne, 2Institute of Earth Sciences, University of Lausanne, CH-1015

Breakthrough curves (BTCs) are efficient tools to study the transport of bacteria in porous media. Here we introduce tools based on fluidic devices in combination with microscopy and flow cytometric counting to obtain BTCs.

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Bioengineering

OnePot PURE Cell-Free System
Laura Grasemann *1, Barbora Lavickova *1, M. Carolina Elizondo-Cantú 1, Sebastian J. Maerkl 1
1Institute of Bioengineering, School of Engineering, École Polytechnique Fédérale de Lausanne

We present a fast and cost-effective method to produce the recombinant PURE cell-free TX-TL system using standard laboratory equipment.

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