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Pasteur Institute

8 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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Immunology and Infection

Imaging InlC Secretion to Investigate Cellular Infection by the Bacterial Pathogen Listeria monocytogenes
Andreas Kühbacher 1,2,3, Edith Gouin 1,2,3, Jason Mercer 4, Mario Emmenlauer 5, Christoph Dehio 5, Pascale Cossart 1,2,3, Javier Pizarro-Cerdá 1,2,3
1Unité des Interactions Bactéries Cellules, Pasteur Institute, 2INSERM U604, 3Institut National de la Recherche Agronomique (INRA), USC2020, 4Institute of Biochemistry, ETH Zürich, 5Focal Area Infection Biology, Biozentrum, University of Basel

Listeria monocytogenes is a Gram positive bacterial pathogen frequently used as a major model for the study of intracellular parasitism. Imaging late L. monocytogenes infection stages within the context of small-interfering RNA screens allows for the global study of cellular pathways required for bacterial infection of target host cells.

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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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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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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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