Tissue-Resident Macrophages

This detailed book delves into the diverse techniques and applications to target, isolate, image, phenotype, and analyze tissue-resident and monocyte-derived macrophages. The contents aim to describe the current knowledge about macrophage development and function which forces the scientific field to move beyond the previously described M1/M2 macrophage paradigm to be able to dissect macrophage functions within their specific niches during health and disease. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Thorough and practical, Tissue-Resident Macrophages: Methods and Protocols provides scientists entering the macrophage field with information and tools that allow them to dive into the state-of-the-art methodology used in this vital field.

Includes cutting-edge techniques Provides step-by-step detail essential for reproducible results Contains key implementation advice from the experts

Inhalt
Macrophage Development and Function.- Fate Mapping Macrophages: From Ontogeny to Functions.- Studying Autophagy in Microglia: Overcoming the Obstacles.- Hemocyte Nuclei Isolation from Adult Drosophila melanogaster for snRNA-Seq.- Isolation of Tissue Macrophages in Adult Zebrafish.- Genetic and Immunohistochemistry Tools to Visualize Rat Macrophages In Situ.- Phenotyping of Macrophages in Human Immune System Mice.- Fate-Mapping of Yolk Sac-Derived Macrophages.- Fate-Mapping of Hematopoietic Stem Cell-Derived Macrophages.- Isolation and Flow Cytometry Analysis of Macrophages from White Adipose Tissue.- Isolation and Flow Cytometry Analysis of Macrophages from the Dermis.- Isolation and Flow Cytometry Analysis of Macrophages from the Kidney.- Isolation and Flow Cytometry Analysis of Intestinal Macrophages.- Isolation and Characterization of Testis Macrophages Using Flow Cytometry.- Studying Macrophages in the Murine Fatty Liver Using Flow Cytometry and Confocal Microscopy.- Isolation, Ex Vivo Expansion, and Lentiviral Transduction of Alveolar Macrophages.- Translatome Profiling of Tissue-Resident Macrophages Using the RiboTag Approach.- Spectral Flow Cytometry Analysis of Resident Tissue Macrophages.- Unveiling Macrophage Heterogeneity and Their Spatial Distribution Using Multiplexed Tissue Imaging.- 3D Imaging of Macrophages in Complete Organs.- Whole-Mount Imaging of Adipose Tissue Macrophages.- Functional In Vivo Imaging of Macrophages.- Elucidating Immune Monitoring of Tissue-Resident Macrophages by Intravital Microscopy.- Combined Host-Pathogen Fate Mapping to Investigate Lung Macrophages in Viral Infection.- Measuring the Metabolic State of Tissue-Resident Macrophages via SCENITH.- Analyzing Fcg-Receptor Interactions on Monocytes with the Proximity Ligation Assay (PLA).- Studying Efferocytosis Dynamics in Tissue-Resident Macrophages Ex Vivo.- Monitoring of Inflammasome Activation of Macrophages and Microglia In Vitro, Part 1: Cell Preparationand Inflammasome Stimulation.- Monitoring of Inflammasome Activation of Macrophages and Microglia In Vitro, Part 2: Assessing Inflammasome Activation.- Detection of G-Quadruplex DNA Structures in Macrophages.- Adaptation of Human iPSC-Derived Macrophages Towards an Alveolar Macrophage-Like Phenotype Post Intra-Pulmonary Transfer into Murine Models.- Tackling Tissue Macrophage Heterogeneity by SplitCre Transgenesis.- Automated Cell Counting of Macrophages In Situ.- Morphometric Analyses of Macrophages.- Combined Analysis of mRNA Expression and Open Chromatin in Microglia.