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Title: Protocol for investigating tertiary lymphoid structures in human and murine fixed tissue sections using Opal™-TSA multiplex immunohistochemistry.
Austin Authors: Quigley, Luke T;Pang, Lokman;Tavancheh, Elnaz;Ernst, Matthias ;Behren, Andreas;Huynh, Jennifer;Da Gama Duarte, Jessica
Affiliation: Department of Medicine, University of Melbourne, Parkville, VIC 3010, Australia.
Olivia Newton-John Cancer Research Institute
Issue Date: 17-Mar-2023
Date: 2023
Publication information: STAR Protocols 2023-03-17; 4(1)
Abstract: 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.
DOI: 10.1016/j.xpro.2022.101961
Journal: STAR Protocols
Start page: 101961
PubMed URL: 36633948
PubMed URL:
ISSN: 2666-1667
Type: Journal Article
Subjects: Cancer
Appears in Collections:Journal articles

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