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

STAR Protocols (Mar 2023)

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

Affiliations

Luke T. Quigley: Olivia Newton-John Cancer Research Institute, and School of Cancer Medicine, La Trobe University, Heidelberg, VIC 3084, Australia
Lokman Pang: Olivia Newton-John Cancer Research Institute, and School of Cancer Medicine, La Trobe University, Heidelberg, VIC 3084, Australia
Elnaz Tavancheh: Olivia Newton-John Cancer Research Institute, and School of Cancer Medicine, La Trobe University, Heidelberg, VIC 3084, Australia
Matthias Ernst: Olivia Newton-John Cancer Research Institute, and School of Cancer Medicine, La Trobe University, Heidelberg, VIC 3084, Australia
Andreas Behren: Olivia Newton-John Cancer Research Institute, and School of Cancer Medicine, La Trobe University, Heidelberg, VIC 3084, Australia; Department of Medicine, University of Melbourne, Parkville, VIC 3010, Australia
Jennifer Huynh: Olivia Newton-John Cancer Research Institute, and School of Cancer Medicine, La Trobe University, Heidelberg, VIC 3084, Australia
Jessica Da Gama Duarte: Olivia Newton-John Cancer Research Institute, and School of Cancer Medicine, La Trobe University, Heidelberg, VIC 3084, Australia; Corresponding author

Journal volume & issue: Vol. 4, no. 1
p. 101961

Abstract

Read online

Summary: 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. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

About the journal

Abstract

Keywords