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Multi-color Flow Cytometry Protocol to Characterize Myeloid Cells in Mouse Retina Research

Myeloid cells, specifically microglia and macrophages, are activated in retinal diseases and can improve or worsen retinopathy outcomes based on their inflammatory phenotype. However, assessing the myeloid cell response after retinal injury in mice remains challenging due to the small tissue size an...

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Autores principales: Xiao, Wei, Shahror, Rami A., Morris, Carol A., Caldwell, Ruth B., Fouda, Abdelrahman Y.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Bio-Protocol 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10450788/
https://www.ncbi.nlm.nih.gov/pubmed/37638294
http://dx.doi.org/10.21769/BioProtoc.4745
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author Xiao, Wei
Shahror, Rami A.
Morris, Carol A.
Caldwell, Ruth B.
Fouda, Abdelrahman Y.
author_facet Xiao, Wei
Shahror, Rami A.
Morris, Carol A.
Caldwell, Ruth B.
Fouda, Abdelrahman Y.
author_sort Xiao, Wei
collection PubMed
description Myeloid cells, specifically microglia and macrophages, are activated in retinal diseases and can improve or worsen retinopathy outcomes based on their inflammatory phenotype. However, assessing the myeloid cell response after retinal injury in mice remains challenging due to the small tissue size and the challenges of distinguishing microglia from infiltrating macrophages. In this protocol paper, we describe a flow cytometry–based protocol to assess retinal microglia/macrophage and their inflammatory phenotype after injury. The protocol is amenable to the incorporation of other markers of interest to other researchers. Key features This protocol describes a flow cytometry–based method to analyze the myeloid cell response in retinopathy mouse models. The protocol can distinguish between microglia- and monocyte-derived macrophages. It can be modified to incorporate markers of interest. We show representative results from three different retinopathy models, namely ischemia-reperfusion injury, endotoxin-induced uveitis, and oxygen-induced retinopathy.
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spelling pubmed-104507882023-08-26 Multi-color Flow Cytometry Protocol to Characterize Myeloid Cells in Mouse Retina Research Xiao, Wei Shahror, Rami A. Morris, Carol A. Caldwell, Ruth B. Fouda, Abdelrahman Y. Bio Protoc Methods Article Myeloid cells, specifically microglia and macrophages, are activated in retinal diseases and can improve or worsen retinopathy outcomes based on their inflammatory phenotype. However, assessing the myeloid cell response after retinal injury in mice remains challenging due to the small tissue size and the challenges of distinguishing microglia from infiltrating macrophages. In this protocol paper, we describe a flow cytometry–based protocol to assess retinal microglia/macrophage and their inflammatory phenotype after injury. The protocol is amenable to the incorporation of other markers of interest to other researchers. Key features This protocol describes a flow cytometry–based method to analyze the myeloid cell response in retinopathy mouse models. The protocol can distinguish between microglia- and monocyte-derived macrophages. It can be modified to incorporate markers of interest. We show representative results from three different retinopathy models, namely ischemia-reperfusion injury, endotoxin-induced uveitis, and oxygen-induced retinopathy. Bio-Protocol 2023-08-20 /pmc/articles/PMC10450788/ /pubmed/37638294 http://dx.doi.org/10.21769/BioProtoc.4745 Text en ©Copyright : © 2023 The Authors; This is an open access article under the CC BY-NC license https://creativecommons.org/licenses/by-nc/4.0/This is an open access article under the CC BY-NC license (https://creativecommons.org/licenses/by-nc/4.0/).
spellingShingle Methods Article
Xiao, Wei
Shahror, Rami A.
Morris, Carol A.
Caldwell, Ruth B.
Fouda, Abdelrahman Y.
Multi-color Flow Cytometry Protocol to Characterize Myeloid Cells in Mouse Retina Research
title Multi-color Flow Cytometry Protocol to Characterize Myeloid Cells in Mouse Retina Research
title_full Multi-color Flow Cytometry Protocol to Characterize Myeloid Cells in Mouse Retina Research
title_fullStr Multi-color Flow Cytometry Protocol to Characterize Myeloid Cells in Mouse Retina Research
title_full_unstemmed Multi-color Flow Cytometry Protocol to Characterize Myeloid Cells in Mouse Retina Research
title_short Multi-color Flow Cytometry Protocol to Characterize Myeloid Cells in Mouse Retina Research
title_sort multi-color flow cytometry protocol to characterize myeloid cells in mouse retina research
topic Methods Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10450788/
https://www.ncbi.nlm.nih.gov/pubmed/37638294
http://dx.doi.org/10.21769/BioProtoc.4745
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