Evaluation of Myelin Phagocytosis by Microglia/Macrophages in Nervous Tissue Using Flow Cytometry

Ariadna Regina Gómez-López, Gemma Manich*, Mireia Recasens, Beatriz Almolda, Berta González, Bernardo Castellano

*Corresponding author for this work

Research output: Contribution to journalArticleResearchpeer-review

1 Citation (Scopus)

Abstract

Determination of microglial phagocytosis of myelin has acquired importance in the study of demyelinating diseases. One strategy to determine microglial phagocytosis capacity consists of assaying microglia with fluorescently labeled myelin; however, most approaches are performed in cell culture, where microglia usually show important phenotypic differences compared with in vivo conditions. In this article we describe an adapted flow cytometry protocol to assay myelin phagocytosis by microglia obtained directly from in vivo tissue of the central nervous system. Key steps for a first analysis of phagocytic microglia are provided. Additionally, we describe how to fluorescently label myelin using a pH-sensitive tag, pHrodo™ Green STP Ester.

Original languageEnglish
Article numbere73
JournalCurrent Protocols
Volume1
Issue number3
DOIs
Publication statusPublished - Mar 2021

Keywords

  • flow cytometry
  • microglia
  • myelin
  • phagocytosis
  • pHrodo

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