Whole-mount immunofluorescence imaging and isolation of mesothelium-bound immune cell aggregates during mouse peritoneal inflammation

Margarita Ferriz; Adrián Vega-Pérez; Alejandra Gutiérrez-González; Natalia Alvarez-Ladrón; Carlos Ardavín

doi:10.1016/j.xpro.2023.102079

Whole-mount immunofluorescence imaging and isolation of mesothelium-bound immune cell aggregates during mouse peritoneal inflammation

STAR Protoc. 2023 Mar 17;4(1):102079. doi: 10.1016/j.xpro.2023.102079. Epub 2023 Jan 29.

Authors

Margarita Ferriz¹, Adrián Vega-Pérez¹, Alejandra Gutiérrez-González¹, Natalia Alvarez-Ladrón¹, Carlos Ardavín²

Affiliations

¹ Departamento de Inmunología y Oncología, Centro Nacional de Biotecnología/CSIC, Darwin 3, 28049 Madrid, Spain.
² Departamento de Inmunología y Oncología, Centro Nacional de Biotecnología/CSIC, Darwin 3, 28049 Madrid, Spain. Electronic address: ardavin@cnb.csic.es.

Abstract

Resident peritoneal macrophages (resMØs) are crucial for repairing peritoneal injuries and controlling infections by forming mesothelium-bound resMØ-aggregates in the peritoneal wall and omentum. Here we present a protocol to analyze these structures in mouse models of peritoneal inflammation. We describe the dissection, fixation, immunofluorescent staining, and mounting of whole peritoneal wall and omentum samples and subsequent confocal microscopy imaging of resMØ-aggregates. We also detail the steps to isolate resMØ-aggregates for additional studies, including flow cytometry and electron-microscopy-based analysis. For complete details on the use and execution of this protocol, please refer to Vega-Pérez et al. (2021).¹.

Keywords: Cell Biology; Cell Isolation; Flow Cytometry/Mass Cytometry; Immunology; Microscopy; Model Organisms.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Disease Models, Animal
Epithelium
Fluorescent Antibody Technique
Inflammation*
Mice
Microscopy, Confocal