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University Paris Diderot, Sorbonne Paris Cité, Cellule Pasteur, Institut Pasteur

3 ARTICLES PUBLISHED IN JoVE

Developmental Biology

Characterization of Thymic Settling Progenitors in the Mouse Embryo Using In Vivo and In Vitro Assays

Cyrille Ramond^1,3, Antonio Bandeira², Claire Berthault³, Pablo Pereira³, Ana Cumano³, Odile Burlen-Defranoux³

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.

Genetics

Single-cell Gene Expression Using Multiplex RT-qPCR to Characterize Heterogeneity of Rare Lymphoid Populations

Thibaut Perchet¹, Sylvestre Chea¹, Milena Hasan², Ana Cumano¹, Rachel Golub¹

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.

Biology

Analysis of Cell Suspensions Isolated from Solid Tissues by Spectral Flow Cytometry

Sandrine Schmutz¹, Mariana Valente^2,3,4,5, Ana Cumano², Sophie Novault¹

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.