Isolation and immunofluorescence staining of Aspergillus fumigatus conidia-containing phagolysosomes

Marie Goldmann; Franziska Schmidt; Irene Kyrmizi; Georgios Chamilos; Axel A Brakhage

doi:10.1016/j.xpro.2021.100328

Isolation and immunofluorescence staining of Aspergillus fumigatus conidia-containing phagolysosomes

STAR Protoc. 2021 Feb 5;2(1):100328. doi: 10.1016/j.xpro.2021.100328. eCollection 2021 Mar 19.

Authors

Marie Goldmann¹, Franziska Schmidt^{1

2}, Irene Kyrmizi³, Georgios Chamilos³, Axel A Brakhage^{1

2}

Affiliations

¹ Department of Molecular and Applied Microbiology, Leibniz Institute for Natural Product Research and Infection Biology - Hans Knöll Institute (HKI), 07745 Jena, Germany.
² Department of Microbiology and Molecular Biology, Institute of Microbiology, Friedrich Schiller University Jena, 07745 Jena, Germany.
³ Department of Medicine, University of Crete, and Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology, Heraklion, Crete 71300, Greece.

Abstract

The analysis of phagolysosomes within professional phagocytic cells is facilitated by their isolation. Here, we optimized a protocol for the isolation of intact phagolysosomes from macrophages infected with the spores of Aspergillus fumigatus. Purified phagolysosomes allow improved immunostaining, e.g., of phagolysosomal membrane proteins, or proteome analysis. For complete details on the use and execution of this protocol, please refer to Schmidt et al. (2020).

Keywords: Cell biology; Cell membrane; Immunology; Protein biochemistry.

Publication types

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

MeSH terms

Animals
Aspergillus fumigatus / metabolism*
Fluorescent Antibody Technique
Macrophages* / metabolism
Macrophages* / microbiology
Mice
Phagosomes* / metabolism
Phagosomes* / microbiology
RAW 264.7 Cells
Spores, Fungal / metabolism*