Flow-cytometry-based protocol to analyze respiratory chain function in mouse microglia

Daniel Erny; Nikolaos Dokalis; Charlotte Mezö; Omar Mossad; Thomas Blank; Marco Prinz

doi:10.1016/j.xpro.2022.101186

Flow-cytometry-based protocol to analyze respiratory chain function in mouse microglia

STAR Protoc. 2022 Feb 18;3(1):101186. doi: 10.1016/j.xpro.2022.101186. eCollection 2022 Mar 18.

Authors

Daniel Erny^{1

2}, Nikolaos Dokalis^{1

3}, Charlotte Mezö^{1

3}, Omar Mossad^{1

3}, Thomas Blank¹, Marco Prinz^{1

4

5}

Affiliations

¹ Institute of Neuropathology, University of Freiburg, Freiburg, Germany.
² Berta-Ottenstein-Programme, Faculty of Medicine, University of Freiburg, Freiburg im Breisgau, Germany.
³ Faculty of Biology, University of Freiburg, Freiburg im Breisgau, Germany.
⁴ Center for Basics in NeuroModulation (NeuroModulBasics), Faculty of Medicine, University of Freiburg, Freiburg, Germany.
⁵ Signalling Research Centres BIOSS and CIBSS, University of Freiburg, Freiburg, Germany.

Abstract

Most of the protocols to analyze metabolic features of cell populations from different tissues rely on in vitro cell culture conditions. Here, we present a flow-cytometry-based protocol for measuring the respiratory chain function in permeabilized mouse microglia ex vivo. We describe microglial cell isolation, followed by analyzing complex I and II using flow cytometry. This optimized protocol requires a low input of permeabilized cells and can be applied to other ex vivo isolated cells or cells derived from cell cultures. For complete details on the use and execution of this protocol, please refer to Erny et al. (2021).

Keywords: Cell Biology; Cell isolation; Flow Cytometry/Mass Cytometry; Immunology; Metabolism; Neuroscience.

Publication types

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

MeSH terms

Animals
Cell Culture Techniques*
Cell Separation / methods
Electron Transport
Flow Cytometry / methods
Mice
Microglia*