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Université de Sherbrooke

16 ARTICLES PUBLISHED IN JoVE

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Biology

Preparation of Rat Tail Tendons for Biomechanical and Mechanobiological Studies
Amélie Bruneau 1, Nadia Champagne 1, Paule Cousineau-Pelletier 1, Gabriel Parent 1, Eve Langelier 1
1Groupe PERSEUS, Faculté de Génie Département de génie mécanique, Université de Sherbrooke

This article describes the experimental procedures used to prepare rat tail tendons for biomechanical and mechanobiological studies. Several features of the main steps in preparation are demonstrated, beginning with extraction, cross-sectional area measurement, rinsing and loading into the bioreactor chamber.

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Engineering

Nanofabrication of Gate-defined GaAs/AlGaAs Lateral Quantum Dots
Chloé Bureau-Oxton 1, Julien Camirand Lemyre 1, Michel Pioro-Ladrière 1
1Département de Physique, Faculté des Sciences, Université de Sherbrooke

This paper presents a detailed fabrication protocol for gate-defined semiconductor lateral quantum dots on gallium arsenide heterostructures. These nanoscale devices are used to trap few electrons for use as quantum bits in quantum information processing or for other mesoscopic experiments such as coherent conductance measurements.

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Neuroscience

A Dual Tracer PET-MRI Protocol for the Quantitative Measure of Regional Brain Energy Substrates Uptake in the Rat
Maggie Roy 1, Scott Nugent 1, Sébastien Tremblay 2, Maxime Descoteaux 3, Jean-François Beaudoin 2, Luc Tremblay 2, Roger Lecomte 2,4, Stephen C Cunnane 1
1Research Center on Aging and Department of Physiology and Biophysics, Université de Sherbrooke, 2Sherbrooke Molecular Imaging Center, Étienne-Le Bel Clinical Research Center, Université de Sherbrooke, 3Department of Computer Science, Université de Sherbrooke, 4Department of Nuclear Medicine and Radiobiology, Université de Sherbrooke

Small-animal positron emission tomography enables the assessment of the brain's two main energy substrates: glucose and ketones. In the present method, 11C-acetoacetate and 18F-fluorodeoxyglucose are injected sequentially in each animal, and their uptake is measured quantitatively in specific brain regions determined from the magnetic resonance images.

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Biology

Simultaneous pH Measurement in Endocytic and Cytosolic Compartments in Living Cells using Confocal Microscopy
Fabrice Lucien 1, Kelly Harper 1, Pierre-Paul Pelletier 1, Leonid Volkov 2, Claire M. Dubois 1
1Department of Pediatrics, Faculty of Medicine and Health Sciences, Université de Sherbrooke, 2Biophotonics Core Facility, Centre de Recherche Clinique Etienne-Le Bel

Several methods are available for measuring intracellular pH but very few of these allow simultaneous measurement of cytoplasmic and organellar pH. Here, we describe in detail a rapid and accurate methodology to simultaneously measure cytoplasmic and vesicular pH by ratiometric imaging of living cells.

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Engineering

Selective Area Modification of Silicon Surface Wettability by Pulsed UV Laser Irradiation in Liquid Environment
Neng Liu 1, Khalid Moumanis 1, Jan J. Dubowski 1
1Laboratory for Quantum Semiconductors and Photon-based BioNanotechnology, Interdisciplinary Institute for Technological Innovation, Laboratoire Nanotechnologies Nanosystèmes (LN2)- CNRS UMI-3463, Faculty of Engineering, Université de Sherbrooke

We report on a process of in situ alteration of HF treated Si (001) surface into a hydrophilic or hydrophobic state by irradiating samples in microfluidic chambers filled with H2O2/H2O solution (0.01%-0.5%) or methanol solutions using pulsed UV laser of a relative low pulse fluence.

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Immunology and Infection

Immunofluorescence to Monitor the Cellular Uptake of Human Lactoferrin and its Associated Antiviral Activity Against the Hepatitis C Virus
Andréa Allaire 1, Frédéric Picard-Jean 1, Martin Bisaillon 1
1Département de biochimie, Faculté de médecine et des sciences de la santé, Université de Sherbrooke

The human lactoferrin (hLF) is a component of the immune system. In this study, immunofluorescence assays are used to demonstrate both the hepatocellular uptake of hLF and a qualitative reduction in the hepatitis C virus replication upon treatment with hLF.

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Developmental Biology

Step-specific Sorting of Mouse Spermatids by Flow Cytometry
Olivier Simard *1, Frédéric Leduc *1, Geneviève Acteau 1, Mélina Arguin 1, Marie-Chantal Grégoire 1, Marc-André Brazeau 1, Isabelle Marois 2, Martin V. Richter 2, Guylain Boissonneault 1
1Department of Biochemistry, Université de Sherbrooke, 2Department of Medicine, Université de Sherbrooke

We describe a sorting strategy for mouse spermatids using flow cytometry. Spermatids are sorted into four highly pure populations, including round (spermiogenesis steps 1-9), early elongating (spermiogenesis steps 10-12), late elongating (spermiogenesis steps 13-14) and elongated spermatids (spermiogenesis steps 15-16). DNA staining, size and granulosity are used as selection parameters.

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JoVE Journal

A Simple and Efficient Protocol for the Catalytic Insertion Polymerization of Functional Norbornenes
Basile Commarieu 1, Jonathan Potier 1, Moubarak Compaore 1, Raphael de Boever 1, Regis Imbeault 1, Jerome P. Claverie 1
1Department of Chemistry, Université de Sherbrooke

We describe the catalytic insertion polymerization of 5-norbornene-2-carboxylic acid and 5-vinyl-2-norbornene to form functional polymers with a very high glass transition temperature.

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Bioengineering

Initial Evaluation of Antibody-conjugates Modified with Viral-derived Peptides for Increasing Cellular Accumulation and Improving Tumor Targeting
Simon Beaudoin 1, Michel Paquette 1, Laurent Fafard-Couture 1, Mylene A. Tremblay 1, Roger Lecomte 1,2,3, Brigitte Guérin 1,2,3, Jeffrey V. Leyton 1,2,3
1Department of Nuclear Medicine and Radiobiology, Université de Sherbrooke, 2Sherbrooke Molecular Imaging Center (CIMS), Université de Sherbrooke, 3Sherbrooke Institute of Pharmacology

Viral-derived peptides coupled to antibody-conjugates (ACs) is an approach gaining momentum due to the potential of delivering molecular payloads with increased tumor cell accumulation. Utilizing common methods to evaluate peptide conjugation, AC and payload intracellular accumulation, and tumor targeting, this protocol helps researchers during the key initial development phases.

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Genetics

Investigation of Protein Recruitment to DNA Lesions Using 405 Nm Laser Micro-irradiation
Antoine Gaudreau-Lapierre *1, Daniel Garneau *1, Billel Djerir *1, Frédéric Coulombe 1, Théo Morin 1, Alexandre Marechal 1
1Department of Biology, Université de Sherbrooke

Studying DNA damage repair kinetics requires a system to induce lesions at defined sub-nuclear regions. We describe a method to create localized double-stranded breaks using a laser-scanning confocal microscope equipped with a 405 nm laser and provide automated procedures to quantify the dynamics of repair factors at these lesions.

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Behavior

Driving Under the Influence: How Music Listening Affects Driving Behaviors
Jordan Navarro 1,2, François Osiurak 1,2, Vivien Gaujoux 1, Marie Claude Ouimet 3, Emanuelle Reynaud 1
1Laboratoire d'Etude des Mécanismes Cognitifs (EA 3082), University Lyon 2, 2Institut Universitaire de France, 3Faculté de médecine et des sciences de la santé, Université de Sherbrooke

Here, we present a protocol to assess driving behaviors while following a vehicle in a simulated driving environment. The presented protocol is used to compare the impact of different auditory backgrounds on driving behaviors.

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Medicine

Assessment of the Efficacy of An Osteopathic Treatment in Infants with Biomechanical Impairments to Suckling
Juliette Herzhaft-Le Roy 1, Marianne Xhignesse 2, Isabelle Gaboury 2
1Entraide Naturo-Lait, 2Faculté de médecine et des sciences de la santé, Université de Sherbrooke

Osteopathy is an emerging field of clinical research. Here we present a protocol to assess the efficacy of an osteopathic intervention coupled with lactation consultation, in infants with biomechanical issues impeding breastfeeding.

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Genetics

Mass Spectrometry-Based Proteomics Analyses Using the OpenProt Database to Unveil Novel Proteins Translated from Non-Canonical Open Reading Frames
Marie A. Brunet 1,2, Xavier Roucou 1,2
1Department of Biochemistry, Université de Sherbrooke, 2PROTEO, Quebec Network for Research on Protein Function, Structure, and Engineering

OpenProt is a freely accessible database that enforces a polycistronic model of eukaryotic genomes. Here, we present a protocol for the use of OpenProt databases when interrogating mass spectrometry datasets. Using OpenProt database for analysis of proteomic experiments allows for discovery of novel and previously undetectable proteins.

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Biochemistry

MS2-Affinity Purification Coupled with RNA Sequencing in Gram-Positive Bacteria
Noémie Mercier 1, Karine Prévost 2, Eric Massé 2, Pascale Romby 1, Isabelle Caldelari 1, David Lalaouna 1
1Université de Strasbourg, CNRS, 2Department of Biochemistry, Université de Sherbrooke

MAPS technology has been developed to scrutinize the targetome of a specific regulatory RNA in vivo. The sRNA of interest is tagged with a MS2 aptamer enabling the co-purification of its RNA partners and their identification by RNA sequencing. This modified protocol is particularly suited for Gram-positive bacteria. 

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Neuroscience

Whole Central and Peripheral Nervous System Mice Dissection
Krystina Rhéaume *1, Zhiguo Chen *2, Yong Wang *2, Camille Plante 1, Dhanushka Hewa Bostanthirige 1, Maude Lévesque 1, Sameh Geha 3,4, Lu Q. Le 2,5,6,7, Jean-Philippe Brosseau 1,4
1Department of Biochemistry and Functional Genomics, Université de Sherbrooke, 2Department of Dermatology, University of Texas Southwestern Medical Center at Dallas, 3Department of Pathology, Université de Sherbrooke, 4Centre de Recherche du Centre Hospitalier de Universitaire de Sherbrooke, Université de Sherbrooke, 5Simmons Comprehensive Cancer Center, University of Texas Southwestern Medical Center at Dallas, 6UTSW Comprehensive Neurofibromatosis Clinic, University of Texas Southwestern Medical Center at Dallas, 7Hamon Center for Regenerative Science and Medicine, University of Texas Southwestern Medical Center at Dallas

Here, we present a protocol to complete the dissection of central and peripheral nervous systems of mice. The dissected tissues are further analyzed in downstream applications, such as taking gross pictures or histology.

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Biology

Growing a Cystic Fibrosis-Relevant Polymicrobial Biofilm to Probe Community Phenotypes
Sarah Poirier 1, Fabrice Jean-Pierre 1
1Département de Biologie, Université de Sherbrooke

This protocol describes a cystic fibrosis (CF) lung-relevant four-species polymicrobial biofilm model that can be used to explore the impact of bacterial interspecies interactions.

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