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Generation of Bone Marrow Derived Murine Dendritic Cells for Use in 2-photon Imaging

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Published on: July 9, 2008

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

Felix Bayerl¹, David A Bejarano², Giulia Bertacchi³

¹Institute of Molecular Immunology, School of Medicine, Klinikum Rechts der Isar, Technical University of Munich (TUM), Ismaninger Str. 22, Munich, Germany.

European Journal of Immunology

|January 9, 2023

Summary

This guide details advanced microscopy techniques for studying dendritic cells (DC). It covers DC preparation, imaging, and functional analysis in various tissues, aiding immunologists in understanding immune responses.

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Area of Science:

Immunology
Cell Biology
Microscopy

Background:

Dendritic cells (DC) are crucial immune cells involved in initiating adaptive immunity.
Understanding DC spatial organization, migration, and communication within tissues is vital for dissecting immune responses.
Existing protocols may not fully capture the dynamic nature and complex functions of DCs in situ.

Purpose of the Study:

To provide state-of-the-art protocols for the comprehensive analysis of mouse and human dendritic cells.
To detail multiparameter fluorescence microscopy methods for exploring DC spatial organization and function in diverse tissue environments.
To enable the study of DC dynamics, T cell cross-talk, and interactions with other tissue cells.

Main Methods:

Preparation and phenotype analysis of dendritic cells (DCs) from lymphoid and non-lymphoid organs.
Multiparameter fluorescence microscopy for visualizing DC subsets and their functions in fixed tissue sections.
Intravital microscopy for studying DC dynamics and T cell interactions in vivo.
3D cell culture models for analyzing DC interactions with tissue cells.

Main Results:

Detailed procedures for advanced imaging techniques are presented.
Methods allow for large-scale visualization, identification, and quantitative analysis of DC subsets.
Protocols facilitate the study of DC migration, communication, and functional roles in immunity.
The study enables exploration of DC interactions with tissue cells and T cells.

Conclusions:

This article serves as an essential resource for basic and clinical immunologists studying dendritic cells.
The provided protocols enhance the understanding of dendritic cell biology and their roles in immune regulation.
Advanced microscopy techniques are critical for dissecting the complex spatial and functional aspects of dendritic cells in immunity.