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Institut Pasteur

29 ARTICLES PUBLISHED IN JoVE

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Advanced Biology

Flow Cytometry and Fluorescence-Activated Cell Sorting (FACS): Isolation of Splenic B Lymphocytes
Perchet Thibaut 1,2,3, Sophie1 Novault 1,2,3, Sophie Novault 4, Rachel Golub 1,2,3
1Unit for Lymphpoiesis, Department of Immunology, Pasteur Institute, 2INSERM U1223, 3Université Paris Diderot, Sorbonne Paris Cité, Cellule Pasteur, 4Flow Cytometry Platfrom, Cytometry and Biomarkers UtechS, Center for Translational Science, Pasteur Institute

Flow Cytometry and Fluorescence-Activated Cell Sorting (FACS): Isolation of Splenic B Lymphocytes

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Advanced Biology

Magnetic Activated Cell Sorting (MACS): Isolation of Thymic T Lymphocytes
Meunier Sylvain 1,2,3, Perchet Thibaut 1,2,3, Sophie Novault 4, Rachel Golub 1,2,3
1Unit for Lymphopoiesis, Department of Immunology, Pasteur Institute, 2INSERM U1223, 3Université Paris Diderot, Sorbonne Paris Cité Cellule Pasteur, 4Flow Cytometry Platfrom, Cytometry and Biomarkers UtechS, Center for Translational Science, Pasteur Institute

Magnetic Activated Cell Sorting (MACS): Isolation of Thymic T Lymphocytes

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Advanced Biology

Cell Cycle Analysis: Assessing CD4 and CD8 T Cell Proliferation After Stimulation Using CFSE Staining and Flow Cytometry
Perchet Thibaut 1,2,3, Meunier Sylvain 1,2,3, Sophie Novault 4, Rachel Golub 1,2,3
1Unit for Lymphopoiesis, Department of Immunology, Pasteur Institute, 2INSERM U1223, 3Université Paris Diderot, Sorbonne Paris Cité Cellule Pasteur, 4Flow Cytometry Platform, Cytometry and Biomarkers UtechS, Center for Translational Science, Pasteur Institute

Cell Cycle Analysis: Assessing CD4 and CD8 T Cell Proliferation After Stimulation Using CFSE Staining and Flow Cytometry

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Advanced Biology

Adoptive Cell Transfer: Introducing Donor Mouse Splenocytes to a Host Mouse and Assessing Success via FACS
Meunier Sylvain 1,2,3, Perchet Thibaut 1,2,3, Sophie Novault 4, Rachel Golub 1,2,3
1Unit for Lymphopoiesis, Department of Immunology, Pasteur Institute, 2INSERM U1223, 3Université Paris Diderot, Sorbonne Paris Cité Cellule Pasteur, 4Flow Cytometry Platfrom, Cytometry and Biomarkers UtechS, Center for Translational Science, Pasteur Institute

Adoptive Cell Transfer: Introducing Donor Mouse Splenocytes to a Host Mouse and Assessing Success via FACS

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JoVE Methods Collections: Current Research Methods in Rabies Diagnosis, Prevention, Treatment, and Control
Charles E. Rupprecht 1, Wei-Cheng Hsu 2, Feng Ye 3, Megan Golding 4, Luca Zaeck 5, Laurent Dacheux 6, Noel Tordo 7, Jodie A. Jarvis 8, Erin Patrick 9, Are Berentsen 10
1LYSSA LLC, 2Animal Health Research Institute, Council of Agriculture, Executive Yuan, 3Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary Medicine, Academy of Military Medical Sciences, 4Wildlife Zoonoses & Vector-Borne Diseases Research Group, Animal and Plant Health Agency, 5Institute of Molecular Virology and Cell Biology, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, 6Unit Lyssavirus Epidemiology and Neuropathology, National Reference Center for Rabies and WHO Collaborating Center for Reference and Research on Rabies, Institut Pasteur, 7Unit Antiviral Strategies, OIE Reference Laboratories for Rift Valley Fever Virus & Crimean Congo Hemorrhagic Fever Virus, Institut Pasteur, Institut Pasteur de Guinea, Conakry, 8Rabies Laboratory, Wadsworth Center, New York State Department of Health, 9Animal and Plant Health Inspection Service, Wildlife Services, Knoxville, TN, US Department of Agriculture, 10Animal and Plant Health Inspection Service, Wildlife Services, National Wildlife Research Center, US Department of Agriculture

JoVE Methods Collections: Current Research Methods in Rabies Diagnosis, Prevention, Treatment, and Control

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Bioengineering

Remote Magnetic Actuation of Micrometric Probes for in situ 3D Mapping of Bacterial Biofilm Physical Properties
Olivier Galy 1, Kais Zrelli 1, Patricia Latour-Lambert 2, Lyndsey Kirwan 3, Nelly Henry 1,3
1Physicochime Curie, CNRS UMR 168, Institut Curie, Sorbonne Universités, UPMC, 2Unité de Génétique des Biofilms, Institut Pasteur, 3Laboratoire Jean Perrin, CNRS UMR 8237, Sorbonne Universités, UPMC

This paper shows an original methodology based on the remote actuation of magnetic particles seeded in a bacterial biofilm and the development of dedicated magnetic tweezers to measure in situ the local mechanical properties of the complex living material built by micro-organisms at interfaces.

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Immunology and Infection

High-throughput Screening for Broad-spectrum Chemical Inhibitors of RNA Viruses
Marianne Lucas-Hourani 1, Hélène Munier-Lehmann 2, Olivier Helynck 2, Anastassia Komarova 1, Philippe Desprès 3, Frédéric Tangy 1, Pierre-Olivier Vidalain 1
1Unité de Génomique Virale et Vaccination, Virology Department, Institut Pasteur, CNRS UMR3569, 2Unité de Chimie et Biocatalyse, Biochemistry and Structural Biology Department, Institut Pasteur, CNRS UMR3523, 3Unité des Interactions Moléculaires Flavivirus-Hôtes, Virology Department, Institut Pasteur

In vitro assays to measure virus replication have been greatly improved by the development of recombinant RNA viruses expressing luciferase or other enzymes capable of bioluminescence. Here we detail a high-throughput screening pipeline that combines such recombinant strains of measles and chikungunya viruses to isolate broad-spectrum antivirals from chemical libraries.

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Bioengineering

A Step Beyond BRET: Fluorescence by Unbound Excitation from Luminescence (FUEL)
Joseph Dragavon 1, Carolyn Sinow 2, Alexandra D. Holland 1, Abdessalem Rekiki 1, Ioanna Theodorou 3, Chelsea Samson 4, Samantha Blazquez 1, Kelly L. Rogers 5, Régis Tournebize 1,6,7, Spencer L. Shorte 1
1Plate-Forme d'Imagerie Dynamique, Imagopole, Institut Pasteur, 2Department of Radiation Oncology, Stanford School of Medicine, 3Service Hospitalier Frédéric Joliot, Institut d'Imagerie Biomédicale, 4Vanderbilt School of Medicine, 5The Walter & Eliza Hall Institute of Medical Research, 6Unité INSERM U786, Institut Pasteur, 7Unité de Pathogénie Microbienne Moléculaire, Institut Pasteur

Expanding the foundation and applicability of Fluorescence by Unbound Excitation from Luminescence (FUEL) by surveying the relevant principles and demonstrating its compatibility with a multitude of fluorophores and antibody-targeted conditions.

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Neuroscience

Using Insect Electroantennogram Sensors on Autonomous Robots for Olfactory Searches
Dominique Martinez 1, Lotfi Arhidi 1, Elodie Demondion 2, Jean-Baptiste Masson 3, Philippe Lucas 2
1UMR 7503, Laboratoire Lorrain de Recherche en Informatique et ses Applications (LORIA), Centre National de la Recherche Scientifique (CNRS), 2UMR 1392 iEES-Paris, Institut d'Ecologie et des Sciences de l'Environnement de Paris, 3Physics of Biological Systems, Institut Pasteur

We describe a protocol for using insect antennae in the form of electroantennograms (EAGs) on autonomous robots. Our experimental design allows stable recordings within a day and resolves individual odor patches up to 10 Hz. The efficiency of EAG sensors for olfactory searches is demonstrated in driving a robot toward an odor source.

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Developmental Biology

Expression of Fluorescent Proteins in Branchiostoma lanceolatum by mRNA Injection into Unfertilized Oocytes
Estelle Hirsinger 1, João Emanuel Carvalho 2, Christine Chevalier 1,3, Georges Lutfalla 4, Jean-François Nicolas 1, Nadine Peyriéras 5, Michael Schubert 2
1Département de Biologie du Développement et Cellules Souches, Institut Pasteur, 2Laboratoire de Biologie du Développement de Villefranche-sur-Mer (UMR7009 CNRS/UPMC Univ Paris 06), Sorbonne Universités, 3Equipe Epigenetic Control of Normal and Pathological Hematopoiesis, Centre de Recherche en Cancérologie de Marseille, 4Unité de Dynamique des Interactions Membranaires Normales et Pathologiques, CNRS UMR5235/DAA/cc107/Université Montpellier II, 5Plateforme BioEmergences IBiSA FBI, CNRS-NED, Institut de Neurobiologie Alfred Fessard

We report here the robust and efficient expression of fluorescent proteins after mRNA injection into unfertilized oocytes of Branchiostoma lanceolatum. The development of the microinjection technique in this basal chordate will pave the way for far-reaching technical innovations in this emerging model system, including in vivo imaging and gene-specific manipulations.

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Developmental Biology

Characterization of Thymic Settling Progenitors in the Mouse Embryo Using In Vivo and In Vitro Assays
Cyrille Ramond 1,3, Antonio Bandeira 2, Claire Berthault 3, Pablo Pereira 3, Ana Cumano 3, Odile Burlen-Defranoux 3
1Research Center Growth and Signaling, INSERM U845, Institut Cochin, 2Unit for Biology of Lymphocyte Populations, Immunology Department, Institut Pasteur and CIMI, Unity of Treg Biology and Therapy, University of Pierre & Marie Curie, 3Unit for Lymphopoiesis, Immunology Department, INSERM U668, University Paris Diderot, Sorbonne Paris Cité, Cellule Pasteur, Institut Pasteur

This article describes in vivo and in vitro methodology to characterize the thymic settling progenitors by the analysis of the kinetics of generation, phenotype and numbers of their T cell progeny.

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Developmental Biology

Three-dimensional Quantification of Dendritic Spines from Pyramidal Neurons Derived from Human Induced Pluripotent Stem Cells
Laura Gouder 1,2,3, Jean-Yves Tinevez 4, Hany Goubran-Botros 1,2,3, Alexandra Benchoua 5, Thomas Bourgeron 1,2,3, Isabelle Cloëz-Tayarani 1,2,3
1Human Genetics and Cognitive Functions, Institut Pasteur, 2CNRS URA 2182 'Genes, synapses and cognition', Institut Pasteur, 3Human Genetics and Cognitive Functions, Université Paris Diderot, Sorbonne Paris Cité, 4Plateforme d' Imagerie Dynamique, Imagopole, CiTech, Institut Pasteur, 5Neuroplasticity and Therapeutics, CECS, I-STEM, AFM, Evry

Dendritic spines of pyramidal neurons are the sites of most excitatory synapses in mammalian brain cortex. This method describes a 3D quantitative analysis of spine morphologies in human cortical pyramidal glutamatergic neurons derived from induced pluripotent stem cells.

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Immunology and Infection

Myeloid Cell Isolation from Mouse Skin and Draining Lymph Node Following Intradermal Immunization with Live Attenuated Plasmodium Sporozoites
Laura Mac-Daniel 1, Matthew R. Buckwalter 2, Pascale Gueirard 1, Robert Ménard 1
1Unité de Biologie et Génétique du Paludisme, Institut Pasteur, 2Unité dImmunobiologie des Cellules Dendritiques, Institut Pasteur

We describe here a protocol for isolating myeloid cells from mouse skin and draining lymph node following intradermal injection of Plasmodium sporozoites. Flow cytometry of collected cells provides a reliable assay to characterize the skin and draining lymph node inflammatory response to the parasite.

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Behavior

Recording Mouse Ultrasonic Vocalizations to Evaluate Social Communication
Allain-Thibeault Ferhat 1, Nicolas Torquet 2, Anne-Marie Le Sourd 1, Fabrice de Chaumont 3, Jean-Christophe Olivo-Marin 3, Philippe Faure 2, Thomas Bourgeron 1, Elodie Ey 1
1Human Genetics and Cognitive Functions, University Paris Diderot, CNRS UMR 3571, Institut Pasteur, 2Neurophysiology and Behavior, University Pierre et Marie Curie Paris 6, CNRS UMR 7102, 3Bio Image Analysis, CNRS URA 2582, Institut Pasteur

Mouse ultrasonic vocalizations are used as proxies to model the genetic bases of vocal communication deficits in mouse models for neuropsychiatric disorders. The present protocol describes three experimental contexts that reliably elicit ultrasonic vocalizations from pups (throughout development) and adult mice (same-sex interactions, male-estrus female interactions).

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Genetics

Single-cell Gene Expression Using Multiplex RT-qPCR to Characterize Heterogeneity of Rare Lymphoid Populations
Thibaut Perchet 1, Sylvestre Chea 1, Milena Hasan 2, Ana Cumano 1, Rachel Golub 1
1Unit for Lymphopoieseis, Immunology Department, INSERM U1223, University Paris Diderot, Sorbonne Paris Cité, Cellule Pasteur, Institut Pasteur, 2Center for Translational Science, Institut Pasteur, INSERM UMS20

This protocol describes how to assess the expression of a large array of genes at the clonal level. Single-cell RT-qPCR produces highly reliable results with a strong sensitivity for hundreds of samples and genes.

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Developmental Biology

A Versatile Mounting Method for Long Term Imaging of Zebrafish Development
Estelle Hirsinger 1,3, Ben Steventon 1,2
1Department of Developmental and Stem Cell Biology, Institut Pasteur, 2Department of Genetics, University of Cambridge, 3IBPS- Laboratoire de Biologie du Developpement (LBD), CNRS, UPMC, UMR 7622, INSERM ERL U1156

Here, we present a versatile mounting method that allows for the long-term time-lapse imaging of the posterior body development of live zebrafish embryos without perturbing normal development.

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Developmental Biology

Identification Of Erythromyeloid Progenitors And Their Progeny In The Mouse Embryo By Flow Cytometry
Lorea Iturri 1,2, Javier Saenz Coronilla 1, Yvan Lallemand 1, Elisa Gomez Perdiguero 1
1Department of Developmental and Stem Cell Biology, CNRS UMR3738, Department of Immunology, Institut Pasteur, 2Cellule Pasteur UPMC, University Pierre et Marie Curie

While infiltrating macrophages are continuously recruited to adult tissues from circulating precursors, resident macrophages seed their tissue during development, where they are maintained without further input from progenitors. The progenitors for resident macrophages were recently identified. Here, we present methods for the genetic fate mapping of the resident macrophage progenitors.

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Immunology and Infection

Immunofluorescence Analysis of Stress Granule Formation After Bacterial Challenge of Mammalian Cells
Pascale Vonaesch 1, Philippe J. Sansonetti 1,2, Pamela Schnupf 3
1Unité de Pathogénie Microbienne Moléculaire, INSERM U786, Institut Pasteur, 2Microbiologie et Maladies Infectieuses, Collège de France, 3Laboratory of Intestinal Immunity, Institut Imagine-INSERM UMR 1163, Université Paris Descartes-Sorbonne Paris Cité

We describe a method for the qualitative and quantitative analysis of stress granule formation in mammalian cells after the cells are challenged with bacteria and a number of different stresses. This protocol can be applied to investigate the cellular stress granule response in a wide range of host-bacterial interactions.

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Biology

Analysis of Cell Suspensions Isolated from Solid Tissues by Spectral Flow Cytometry
Sandrine Schmutz 1, Mariana Valente 2,3,4,5, Ana Cumano 2, Sophie Novault 1
1Flow Cytometry Core Facility, Center for Translational Research-Technical Core, Institut Pasteur, 2Unit for Lymphopoiesis, Immunology Department, INSERM U1223, University Paris Diderot, Sorbonne Paris Cité, Cellule Pasteur, Institut Pasteur, 3Stem-Cell Microenvironments in Repair/Regeneration Team, Instituto de Investigação e Inovação em Saúde (i3s), INEB - Instituto de Engenharia Biomédica, 4ICBAS - Instituto de Ciências Biomédicas Abel Salazar, Universidade do Porto, 5Stem Cells and Regenerative Medicine Team, UMRS 1166, ICAN - Institute of Cardiometabolism And Nutrition, UPMC - Université Pierre et Marie Curie - Paris 6, INSERM

This article describes spectral cytometry, a new approach in flow cytometry that uses the shapes of emission spectra to distinguish fluorochromes. An algorithm replaces compensations and can treat auto-fluorescence as an independent parameter. This new approach allows for the proper analysis of cells isolated from solid organs.

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Developmental Biology

Evaluation of Injury-induced Senescence and In Vivo Reprogramming in the Skeletal Muscle
Coralie Cazin 1, Aurelie Chiche 1, Han Li 1
1Cellular Plasticity & Disease Modelling, Department of Developmental & Stem Cell Biology, CNRS UMR 3738, Institut Pasteur

Here we present a detailed protocol to detect both senescent and pluripotent stem cells in the skeletal muscle upon injury while inducing in vivo reprogramming. This method is suitable for evaluating the role of cellular senescence during tissue regeneration and reprogramming in vivo.

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Biology

Imaging Intermediate Filaments and Microtubules with 2-dimensional Direct Stochastic Optical Reconstruction Microscopy
Cécile Leduc 1, Audrey Salles 2, Spencer L. Shorte 2, Sandrine Etienne-Manneville 1
1Cell Polarity, Migration and Cancer Unit, UMR 3691, CNRS, Institut Pasteur, 2UTechS Photonic BioImaging (Imagopole) Citech, Institut Pasteur

The overall goal of this methodology is to give the optimal experimental conditions from sample preparation to image acquisition and reconstruction in order to perform 2D dual color dSTORM images of microtubules and intermediate filaments in fixed cells

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Immunology and Infection

Development and Validation of an Ultrasensitive Single Molecule Array Digital Enzyme-linked Immunosorbent Assay for Human Interferon-α
Alba Llibre *1,2, Vincent Bondet *1,2, Mathieu P. Rodero 3, David Hunt 4, Yanick J. Crow 3,5, Darragh Duffy 1,2
1Immunobiology of Dendritic Cells, Institut Pasteur, 2INSERM U1223, 3Laboratory of Neurogenetics and Neuroinflammation, INSERM UMR1163, Institut Imagine, 4MRC Human Genetics Unit, MRC Institute of Genetics and Molecular Medicine, University of Edinburgh, 5Manchester Centre for Genomic Medicine, University of Manchester

Here we present a protocol to describe the development and validation of a single molecule array digital ELISA assay, which enables the ultra-sensitive detection of all IFN-α subtypes in human samples.

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Cancer Research

Selecting Multiple Biomarker Subsets with Similarly Effective Binary Classification Performances
Xin Feng 1, Shaofei Wang 1, Quewang Liu 1, Han Li 2, Jiamei Liu 2, Cheng Xu 2, Weifeng Yang 2, Yayun Shu 2, Weiwei Zheng 1, Bingxin Yu 3, Mingran Qi 4, Wenyang Zhou 1, Fengfeng Zhou 1
1College of Computer Science and Technology, and Key Laboratory of Symbolic Computation and Knowledge Engineering of Ministry of Education, Jilin University, 2College of Software, Jilin University, 3Ultrasonography Department, China-Japan Union Hospital of Jilin University, 4Department of Pathogenobiology, College of Basic Medical Science, Jilin University

Existing algorithms generate one solution for a biomarker detection dataset. This protocol demonstrates the existence of multiple similarly effective solutions and presents a user-friendly software to help biomedical researchers investigate their datasets for the proposed challenge. Computer scientists may also provide this feature in their biomarker detection algorithms.

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Biology

The MUB40 Peptide for Use in Detecting Neutrophil-Mediated Inflammation Events
Mark C. Anderson 1,2, Louise Injarabian 1,2,3, Antonin Andre 1,2,3, Regis Tournebize 1,2,4, Benoit S. Marteyn 1,2
1Unité de Pathogénie Microbienne Moléculaire, Institut Pasteur, 2INSERM Unité 1202, Institut Pasteur, 3Institut de Biochimie et Génétique Cellulaires, Université Bordeaux Segalen, 4Unit of Technology and Service Photonic BioImaging, Institut Pasteur

Here, we present a protocol to detect the presence of neutrophils in fixed/permeabilized histology sections and assess the activation state of live purified neutrophils. In particular, the MUB40 peptide binds lactoferrin present in neutrophil-specific and tertiary granules. Exposure of the granule contents through either permeabilization or neutrophil activation allows for the marking of neutrophils.

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Immunology and Infection

In Vitro ELISA Test to Evaluate Rabies Vaccine Potency
Corinne Jallet 1, Noël Tordo 1,2
1Unit Antiviral Strategies, OIE Reference Laboratories for Rift Valley Fever Virus & Crimean Congo Hemorrhagic Fever Virus, Institut Pasteur, 2Institut Pasteur de Guinée, Conakry

Here we describe an indirect ELISA sandwich immunocapture to determine the immunogenic glycoprotein contents in rabies vaccines. This test uses a neutralizing Monoclonal Antibody (mAb-D1) recognizing glycoprotein trimers. It is an alternative to the in vivo NIH test to follow the consistency of vaccine potency during production.

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Immunology and Infection

Field Postmortem Rabies Rapid Immunochromatographic Diagnostic Test for Resource-Limited Settings with Further Molecular Applications
Stephanie Mauti 1, Monique Léchenne 2, Service Naïssengar 3, Abdallah Traoré 4, Vessaly Kallo 5,6, Casimir Kouakou 7, Emmanuel Couacy-Hymann 7, Morgane Gourlaouen 8, Céline Mbilo 9,10, Pati Patient Pyana 11, Enos Madaye 3, Ibrahima Dicko 4, Pascal Cozette 1, Paola De Benedictis 8, Hervé Bourhy 1, Jakob Zinsstag 9,10, Laurent Dacheux 1
1Unit Lyssavirus Epidemiology and Neuropathology, National Reference Center for Rabies and WHO Collaborating Center for Reference and Research on Rabies, Institut Pasteur, 2Environment and Sustainability Institute, University of Exeter, Penryn Campus, 3Institut de Recherche en Elevage pour le Développement, 4Laboratoire Central Vétérinaire, 5Direction des Services Vétérinaires, 6Ecole Inter Etats de Sciences et de Médecine Vétérinaires de Dakar, 7Laboratoire Central Vétérinaire de Bingerville, Laboratoire National d'Appui au Développement Agricole Bingerville, 8FAO Reference Centre for Rabies, Istituto Zooprofilattico Sperimentale delle Venezie, 9Swiss Tropical and Public Health Institute, 10University of Basel, 11Institut National de Recherche Biomédicale

We present a complete protocol for postmortem diagnosis of animal rabies under field conditions using a rapid immunochromatographic diagnostic test (RIDT), from brain biopsy sampling to final interpretation. We also describe further applications using the device for molecular analysis and viral genotyping.

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Biology

Analyzing Oxidative Stress in Murine Intestinal Organoids using Reactive Oxygen Species-Sensitive Fluorogenic Probe
Aline Stedman 1,3, Antonin Levy 1,4, Philippe J. Sansonetti 1,2,5, Giulia Nigro 1,6
1Molecular Microbial Pathogenesis Unit, Institut Pasteur, 2Chaire de Microbiologie et Maladies Infectieuses, Collège de France, 3Institut de Biologie Paris Seine (IBPS) - Developmental Biology Unit, Sorbonne Université, CNRS UMR7622, INSERM U1156, 4Molecular Radiotherapy, INSERM U1030, Gustave Roussy, Université Paris-Saclay, 5The Center for Microbes, Development and Health, Institut Pasteur Shanghai and Chinese Academy of Sciences, 6Microenvironment and Immunity Unit, Institut Pasteur

The present protocol describes a method to detect reactive oxygen species (ROS) in the intestinal murine organoids using qualitative imaging and quantitative cytometry assays. This work can be potentially extended to other fluorescent probes to test the effect of selected compounds on ROS.

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Biology

Isolation and Characterization of the Immune Cells from Micro-dissected Mouse Choroid Plexuses
Amaia Dominguez-Belloso 1, Sandrine Schmutz 2, Sophie Novault 2, Laetitia Travier *1, Aleksandra Deczkowska *1
1Brain-Immune Communication Lab, Institut Pasteur, 2Cytometry platform, CB UTechS, Institut Pasteur

This study uses flow cytometry and two different gating strategies on isolated perfused mice brain choroid plexuses; this protocol identifies the main immune cell subsets that populate this brain structure.

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Biology

Live Imaging of Microtubule Dynamics in Glioblastoma Cells Invading the Zebrafish Brain
Florent Peglion 1, Franck Coumailleau 1, Sandrine Etienne-Manneville 1
1Cell Polarity, Migration and Cancer Unit, Université Paris-Cité, Institut Pasteur, Équipe Labellisée Ligue Contre le Cancer

We report a technique permitting live imaging of microtubule dynamics in glioblastoma (GBM) cells invading a vertebrate brain tissue. Coupling the orthotopic injection of fluorescently tagged GBM cells into a transparent zebrafish brain with high-resolution intravital imaging allows the measurement of cytoskeleton dynamics during in situ cancer invasion.

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