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Université de Lille

6 ARTICLES PUBLISHED IN JoVE

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Immunology and Infection

A Microscopic Phenotypic Assay for the Quantification of Intracellular Mycobacteria Adapted for High-throughput/High-content Screening
Christophe. J Queval *1, Ok-Ryul Song *1, Vincent Delorme *1, Raffaella Iantomasi 1, Romain Veyron-Churlet 1, Nathalie Deboosère 1, Valérie Landry 1, Alain Baulard 1, Priscille Brodin 1
1Inserm U1019 - CNRS UMR 8024, Institut Pasteur de Lille, Université de Lille

Here, we describe a phenotypic assay applicable to the High-throughput/High-content screens of small-interfering synthetic RNA (siRNA), chemical compound, and Mycobacterium tuberculosis mutant libraries. This method relies on the detection of fluorescently labeled Mycobacterium tuberculosis within fluorescently labeled host cell using automated confocal microscopy.

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Cancer Research

A Detailed Protocol for Characterizing the Murine C1498 Cell Line and its Associated Leukemia Mouse Model
Alexia Mopin 1, Virginie Driss 1, Carine Brinster 1,2
1Institut pour la Recherche sur le Cancer de Lille (IRCL), INSERM, UMR-S-1172, Centre de Recherche Jean-Pierre Aubert (JPARC), 2Université de Lille

This manuscript provides a technical procedure that can be used to characterize C1498 cell cultures in vitro and the acute leukemia induced in mice after their injection. Phenotypic and functional analyses are performed using flow cytometry, immunofluorescence microscopy, cytochemistry and May-Grünwald Giemsa staining.

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Environment

Extraction and Analysis of Microbial Phospholipid Fatty Acids in Soils
Sylvie A. Quideau 1, Anne C.S. McIntosh 2, Charlotte E. Norris 1, Emily Lloret 3, Mathew J.B. Swallow 4, Kirsten Hannam 5
1Department of Renewable Resources, University of Alberta, 2Department of Science, Augustana Faculty, University of Alberta, 3Laboratoire Génie Civil et géo-Environnement, Université de Lille, 4Department of Earth and Environmental Sciences, Mount Royal University, 5Forest Ecology & Production, Great Lakes Forestry Centre, Natural Resources Canada

Phospholipid fatty acids provide information about the structure of soil microbial communities. We present methods for extraction from soil samples with a single-phase chloroform mixture, fractionation of extracted lipids using solid phase extraction columns, and methanolysis to produce fatty acid methyl esters, which are analyzed by capillary gas chromatography.

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Chemistry

Visualizing Lignification Dynamics in Plants with Click Chemistry: Dual Labeling is BLISS!
Clémence Simon 1, Corentin Spriet 1, Simon Hawkins 1, Cedric Lion 1
1UGSF – Unité de Glycobiologie Structurale et Fonctionnelle, CNRS, UMR 8576, Université de Lille

BLISS, a dual labeling protocol for studying lignification dynamics, was developed. Using synthetic monolignol reporters and a sequential combination of SPAAC and CuAAC bioorthogonal click reactions, this methodology paves the way to in-depth analysis of the factors that regulate the biogenesis of lignins in planta.

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Genetics

High-Throughput DNA Plasmid Multiplexing and Transfection Using Acoustic Nanodispensing Technology
Béatrice Colin 1, Nicolas Rocq 2, Benoit Deprez 1, Cyril Couturier 1
1Institut Pasteur de Lille, Institut National de la Santé et de la Recherche Médicale (Inserm) U1177, Drugs and Molecules for Living Systems, Université de Lille, 2L'atelier du Développeur, Marquette lez Lille

This protocol describes high-throughput plasmid transfection of mammalian cells in a 384-well plate using acoustic droplet ejection technology. The time-consuming, error-prone DNA dispensing and multiplexing, but also the transfection reagent dispensing, are software-driven and performed by a nanodispenser device. The cells are then seeded in these prefilled wells.

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Biochemistry

Measuring Interactions between Fluorescent Probes and Lignin in Plant Sections by sFLIM Based on Native Autofluorescence
Christine Terryn 1, Anouck Habrant 2, Gabriel Paës 2, Corentin Spriet 3
1PICT Platform, Université de Reims Champagne-Ardenne, 2Fractionation of AgroResources and Environment (FARE) Laboratory, INRA, Université de Reims Champagne-Ardenne, 3Unité de Glycobiologie Structurale et Fonctionnelle, Université de Lille

This protocol describes an original setup combining spectral and fluorescence lifetime measurements to evaluate Förster resonance energy transfer (FRET) between rhodamine-based fluorescent probes and lignin polymer directly in thick plant sections.

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