Protocol for investigating tertiary lymphoid structures in human and murine fixed tissue sections using Opal™-TSA multiplex immunohistochemistry

Tertiary lymphoid structures (TLSs) are ectopic lymphoid aggregates that can develop within or adjacent to tumors, but protocols that can accurately identify and characterize TLSs are lacking. Here, we present a protocol for the in situ interrogation and characterization of TLSs in human and murine...

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Autores principales: Quigley, Luke T., Pang, Lokman, Tavancheh, Elnaz, Ernst, Matthias, Behren, Andreas, Huynh, Jennifer, Da Gama Duarte, Jessica
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9843255/
https://www.ncbi.nlm.nih.gov/pubmed/36633948
http://dx.doi.org/10.1016/j.xpro.2022.101961
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author Quigley, Luke T.
Pang, Lokman
Tavancheh, Elnaz
Ernst, Matthias
Behren, Andreas
Huynh, Jennifer
Da Gama Duarte, Jessica
author_facet Quigley, Luke T.
Pang, Lokman
Tavancheh, Elnaz
Ernst, Matthias
Behren, Andreas
Huynh, Jennifer
Da Gama Duarte, Jessica
author_sort Quigley, Luke T.
collection PubMed
description Tertiary lymphoid structures (TLSs) are ectopic lymphoid aggregates that can develop within or adjacent to tumors, but protocols that can accurately identify and characterize TLSs are lacking. Here, we present a protocol for the in situ interrogation and characterization of TLSs in human and murine tissue sections using Opal™-tyramide signal amplification multiplex immunohistochemistry. This protocol enables simultaneous detection of up to 7 markers (6 antigens and a DAPI counterstain). We also describe a grading system to identify immature and mature TLSs.
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spelling pubmed-98432552023-01-18 Protocol for investigating tertiary lymphoid structures in human and murine fixed tissue sections using Opal™-TSA multiplex immunohistochemistry Quigley, Luke T. Pang, Lokman Tavancheh, Elnaz Ernst, Matthias Behren, Andreas Huynh, Jennifer Da Gama Duarte, Jessica STAR Protoc Protocol Tertiary lymphoid structures (TLSs) are ectopic lymphoid aggregates that can develop within or adjacent to tumors, but protocols that can accurately identify and characterize TLSs are lacking. Here, we present a protocol for the in situ interrogation and characterization of TLSs in human and murine tissue sections using Opal™-tyramide signal amplification multiplex immunohistochemistry. This protocol enables simultaneous detection of up to 7 markers (6 antigens and a DAPI counterstain). We also describe a grading system to identify immature and mature TLSs. Elsevier 2023-01-10 /pmc/articles/PMC9843255/ /pubmed/36633948 http://dx.doi.org/10.1016/j.xpro.2022.101961 Text en © 2022 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
Quigley, Luke T.
Pang, Lokman
Tavancheh, Elnaz
Ernst, Matthias
Behren, Andreas
Huynh, Jennifer
Da Gama Duarte, Jessica
Protocol for investigating tertiary lymphoid structures in human and murine fixed tissue sections using Opal™-TSA multiplex immunohistochemistry
title Protocol for investigating tertiary lymphoid structures in human and murine fixed tissue sections using Opal™-TSA multiplex immunohistochemistry
title_full Protocol for investigating tertiary lymphoid structures in human and murine fixed tissue sections using Opal™-TSA multiplex immunohistochemistry
title_fullStr Protocol for investigating tertiary lymphoid structures in human and murine fixed tissue sections using Opal™-TSA multiplex immunohistochemistry
title_full_unstemmed Protocol for investigating tertiary lymphoid structures in human and murine fixed tissue sections using Opal™-TSA multiplex immunohistochemistry
title_short Protocol for investigating tertiary lymphoid structures in human and murine fixed tissue sections using Opal™-TSA multiplex immunohistochemistry
title_sort protocol for investigating tertiary lymphoid structures in human and murine fixed tissue sections using opal™-tsa multiplex immunohistochemistry
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9843255/
https://www.ncbi.nlm.nih.gov/pubmed/36633948
http://dx.doi.org/10.1016/j.xpro.2022.101961
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