Flow cytometry protocol to quantify immune cells in the separated epithelium and stroma of herpes simplex virus-1-infected mouse cornea

Existing flow cytometry approaches identify immune cells using the whole infected/inflamed cornea, which limits its ability to distinguish the immune cells infiltrating the corneal epithelium from the corneal stroma. Here, we present a protocol to analyze immune cells in the separated epithelium and...

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Detalles Bibliográficos
Autores principales: Setia, Mizumi, Suvas, Pratima Krishna, Suvas, Susmit
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9898781/
https://www.ncbi.nlm.nih.gov/pubmed/36790766
http://dx.doi.org/10.1016/j.xpro.2023.102056
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author Setia, Mizumi
Suvas, Pratima Krishna
Suvas, Susmit
author_facet Setia, Mizumi
Suvas, Pratima Krishna
Suvas, Susmit
author_sort Setia, Mizumi
collection PubMed
description Existing flow cytometry approaches identify immune cells using the whole infected/inflamed cornea, which limits its ability to distinguish the immune cells infiltrating the corneal epithelium from the corneal stroma. Here, we present a protocol to analyze immune cells in the separated epithelium and stroma from naïve and herpes simplex virus-1 (HSV-1)-infected mouse corneas. We describe steps for viral infection, separation of corneal epithelium from stroma, preparation of a single-cell suspension of the individual epithelium and stroma, and flow cytometry assay.
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spelling pubmed-98987812023-02-05 Flow cytometry protocol to quantify immune cells in the separated epithelium and stroma of herpes simplex virus-1-infected mouse cornea Setia, Mizumi Suvas, Pratima Krishna Suvas, Susmit STAR Protoc Protocol Existing flow cytometry approaches identify immune cells using the whole infected/inflamed cornea, which limits its ability to distinguish the immune cells infiltrating the corneal epithelium from the corneal stroma. Here, we present a protocol to analyze immune cells in the separated epithelium and stroma from naïve and herpes simplex virus-1 (HSV-1)-infected mouse corneas. We describe steps for viral infection, separation of corneal epithelium from stroma, preparation of a single-cell suspension of the individual epithelium and stroma, and flow cytometry assay. Elsevier 2023-01-25 /pmc/articles/PMC9898781/ /pubmed/36790766 http://dx.doi.org/10.1016/j.xpro.2023.102056 Text en © 2023 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Setia, Mizumi
Suvas, Pratima Krishna
Suvas, Susmit
Flow cytometry protocol to quantify immune cells in the separated epithelium and stroma of herpes simplex virus-1-infected mouse cornea
title Flow cytometry protocol to quantify immune cells in the separated epithelium and stroma of herpes simplex virus-1-infected mouse cornea
title_full Flow cytometry protocol to quantify immune cells in the separated epithelium and stroma of herpes simplex virus-1-infected mouse cornea
title_fullStr Flow cytometry protocol to quantify immune cells in the separated epithelium and stroma of herpes simplex virus-1-infected mouse cornea
title_full_unstemmed Flow cytometry protocol to quantify immune cells in the separated epithelium and stroma of herpes simplex virus-1-infected mouse cornea
title_short Flow cytometry protocol to quantify immune cells in the separated epithelium and stroma of herpes simplex virus-1-infected mouse cornea
title_sort flow cytometry protocol to quantify immune cells in the separated epithelium and stroma of herpes simplex virus-1-infected mouse cornea
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9898781/
https://www.ncbi.nlm.nih.gov/pubmed/36790766
http://dx.doi.org/10.1016/j.xpro.2023.102056
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