Guidelines for visualization and analysis of DC in tissues using multiparameter fluorescence microscopy imaging methods

Felix Bayerl (1) , David A Bejarano (2) , Giulia Bertacchi (3) , Anne‐claire Doffin (4) , Elisa Gobbini (4) , Margaux Hubert (4) , Lijian Li (5) , Philippa Meiser (1) , Anna‐marie Pedde (1) , Wilfried Posch (3) , Luise Rupp (6) , Andreas Schlitzer (2) , Marc Schmitz (6) , Barbara U Schraml (7) , Stefan Uderhardt (5) , Jenny Valladeau-Guilemond (4, 8, 9) , Doris Wilflingseder (3) , Viktoria Zaderer (3) , Jan P Böttcher (1)

1 TUM - Technische Universität Munchen - Technical University Munich - Université Technique de Munich
2 Universität Bonn = University of Bonn
3 Leopold Franzens Universität Innsbruck - University of Innsbruck
4 UNICANCER/CRCL - Centre de Recherche en Cancérologie de Lyon
5 FAU - Friedrich-Alexander Universität Erlangen-Nürnberg = University of Erlangen-Nuremberg
6 TU Dresden - Technische Universität Dresden = Dresden University of Technology
7 LMU - Ludwig Maximilian University [Munich] = Ludwig Maximilians Universität München
8 HCL - Hospices Civils de Lyon
9 Centre Léon Bérard [Lyon]

This article is part of the Dendritic Cell Guidelines article series, which provides a collection of state-of-the-art protocols for the preparation, phenotype analysis by flow cytometry, generation, fluorescence microscopy, and functional characterization of mouse and human dendritic cells (DC) from lymphoid organs and various non-lymphoid tissues. Here, we provide detailed procedures for a variety of multiparameter fluorescence microscopy imaging methods to explore the spatial organization of DC in tissues and to dissect how DC migrate, communicate, and mediate their multiple functional roles in immunity in a variety of tissue settings. The protocols presented here entail approaches to study DC dynamics and T cell cross-talk by intravital microscopy, large-scale visualization, identification, and quantitative analysis of DC subsets and their functions by multiparameter fluorescence microscopy of fixed tissue sections, and an approach to study DC interactions with tissue cells in a 3D cell culture model. While all protocols were

Domaines: Immunité adaptative; Imagerie

Felix Bayerl, David A Bejarano, Giulia Bertacchi, Anne‐claire Doffin, Elisa Gobbini, et al.. Guidelines for visualization and analysis of DC in tissues using multiparameter fluorescence microscopy imaging methods. European Journal of Immunology, 2023, 53 (11), pp.2249923. ⟨10.1002/eji.202249923⟩. ⟨hal-04477856⟩