Protocol for investigating tertiary lymphoid structures in human and murine fixed tissue sections using Opal™-TSA multiplex immunohistochemistry
Luke T. Quigley, Lokman Pang, Elnaz Tavancheh, Matthias Ernst, Andreas Behren, Jennifer Huynh, Jessica Da Gama Duarte
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.
Topics & Concepts
MultiplexImmunohistochemistryIn situ hybridizationBiologyLymphatic systemIn situPathologyMolecular biologyChemistryImmunologyMedicineGeneGeneticsGene expressionOrganic chemistryLymphoma Diagnosis and TreatmentCancer Immunotherapy and BiomarkersSingle-cell and spatial transcriptomics