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

doi:10.1002/cpz1.73

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

Department of Cellular Biology, Physiology and Immunology

Research output: Contribution to journal › Article › Research › peer-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 language	English
Article number	e73
Journal	Current Protocols
Volume	1
Issue number	3
DOIs	https://doi.org/10.1002/cpz1.73
Publication status	Published - Mar 2021

Keywords

flow cytometry
microglia
myelin
phagocytosis
pHrodo

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10.1002/cpz1.73

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title = "Evaluation of Myelin Phagocytosis by Microglia/Macrophages in Nervous Tissue Using Flow Cytometry",

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{\texttrademark} Green STP Ester.",

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T1 - Evaluation of Myelin Phagocytosis by Microglia/Macrophages in Nervous Tissue Using Flow Cytometry

AU - Gómez-López, Ariadna Regina

AU - Manich, Gemma

AU - Recasens, Mireia

AU - Almolda, Beatriz

AU - González, Berta

AU - Castellano, Bernardo

PY - 2021/3

Y1 - 2021/3

N2 - 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.

AB - 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.

KW - flow cytometry

KW - microglia

KW - myelin

KW - phagocytosis

KW - pHrodo

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DO - 10.1002/cpz1.73

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VL - 1

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M1 - e73

ER -

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

Abstract

Keywords

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